NOTE. In contrast to proviral DNA, viral RNA may be present in multiple copies in infected cells. The purpose of this report is to describe the
|
|
- Dina Washington
- 5 years ago
- Views:
Transcription
1 JOURNAL OF VIROLOGY, May 1979, p X/79/05/ $02.00/0 Vol. 30, No. 2 NOTE Detection of Virus-Specific RNA in Simian Sarcoma- Leukemia Virus-Infected Cells by In Situ Hybridization to Viral Complementary DNA STEVEN L. KAUFMAN,t ROBERT C. GALLO,`* AND NANCY R. MILLER2 Laboratory of Tumor Cell Biology, National Cancer Institute, Bethesda, Maryland 20014,1 and Bethesda Research Laboratories, Rockville, Maryland Received for publication 4 January 1979 An in situ molecular hybridization system which will detect retrovirus RNA in the cytoplasm of individual virus-infected cells has been developed. The technique was applied to cells infected with simian sarcoma-leukemia virus, where the virusspecific RNA was detected by hybridization to simian sarcoma-leukemia virus 3H-labeled complementary DNA. The system is useful for detecting viral RNAcontaining cells in the presence of an excess of virus-negative cells and for determining which type of cell in a heterogenous population is expressing viral RNA. Cytological or in situ hybridization has been used extensively to find the chromosomal location of DNA sequences complementary to a specific purified cellular RNA (for reviews, see Pardue and Gall [11], Hennig [4], Jones [5], and Wimber and Steffensen [15]; for a quantitative analysis of the system, see Szabo et al. [14]). The technique has also been applied to the analysis of cells infected with DNA viruses. It has been used to detect virus-specific DNA in adenovirus-transformed (9) and in polyomatransformed (10) cells by hybridization of the DNA to complementary viral RNA. Complementary DNA (cdna) from simian virus 40 has been used to localize related sequences in host cell chromosomes (13). The use of in situ hybridization in the detection of viral nucleic acids in cells infected by retroviruses has been limited, however. Loni and Green (7) used viral cdna and RNA to detect viral DNA sequences in nuclei of virus-producing cells transformed by Harvey and Moloney strains of murine sarcoma virus. Haase et al. (2) have used the technique to detect proviral DNA in cells infected with visna virus. In many cells infected with RNA type C viruses, however, the proviral sequences can be present in the cellular DNA at one copy per cell or less. This makes detection difficult, requiring very low backgrounds and quantitation of grains. t Present address: Medical College of Virginia, Richmond, VA In contrast to proviral DNA, viral RNA may be present in multiple copies in infected cells. The purpose of this report is to describe the development of the technique of in situ hybridization to detect viral RNA in type C virusproducing cells. We have used a modification of the technique developed by Harrison et al. (3) who used in situ hybridization to detect globin mrna in fetal mouse liver cells and in Friend virus-transformed cells treated with dimethyl sulfoxide. A similar technique has also been applied in DNA virus systems by Moar and Jones (9) who detected virus-specific RNA in the cytoplasm of adenovirus type 2-transformed rat cells and by McDougall in a recent demonstration of viral RNA in herpes type 2-infected cells (8) and evidence for such sequences in RNA from several human cervical carcinomas (8a). Our results indicate that in situ hybridization can be used to detect RNA of the woolly monkey (simian sarcoma-leukemia) virus (SiSV/SiSAV), an infectious primate type C RNA tumor virus, in the cytoplasm of infected cells which produce the virus. To prepare cytological slides for in situ hybridization, cells grown in tissue culture were trypsinized, the trypsin was neutralized by media containing 10% fetal calf serum, and cells were washed twice with serum-free medium. The cell concentration was adjusted to 106 cells per ml. Portions (200,u) of this suspension were then pipetted into cytocentrifuge buckets and 637
2 638 NOTE centrifuged against microscope slides in a Shandon Cytospin cytocentrifuge at 500 rpm for 5 min. After air drying for a minimum of 15 min, the cells were fixed on the slides in ethanolacetic acid (3:1) at room temperature for 10 min. The slides were air dried and stored at -70 C. Just before hybridization, the fixed slides were treated with 0.2 N HCl for 20 min at room temperature, rinsed briefly in water twice, dehydrated through three subsequent 5-min, room temperature washes of 70, 70, and 100% EtOH, and air dried. For the hybridization, [3H]cDNA with a 103-fold excess of trna was dissolved in 40% formamide-3x SSC (lx SSC = 0.15 M NaCl M sodium citrate) and 2,ul (10,000 cpm, approximately 1 ng of [3H]cDNA) was applied to the spot containing cells. The liquid was covered with a 12-mm-diameter glass cover slip, and the slides were placed in a rack in a light mineral oil bath at 44 C. It is not necessary to seal the cover slips in any manner. Hybridization was carried out for the desired length of time (usually 40 to 44 h, which corresponds to a Cot with respect to cdna of approximately 0.2). The slides were removed from the oil bath, rinsed twice (2 min) in chloroform to remove the oil, and then rinsed three times (5 min) in 2x SSC. The cover slips fall off during the first rinse. The hybrids were heated at 550C in 2x SSC for 1 h to dissociate nonspecific complexes, and then washed in 2x SSC at 4 C for 24 h. The slides were dehydrated through three successive 5-min washes in 70% EtOH, 70% EtOH, and 100% EtOH and were air dried. The presence of a hybrid between the [3H]- cdna and cytoplasmic RNA was detected by autoradiographic exposure to a photographic emulsion which was used to coat the slides. Autoradiography was carried out essentially as described by Pardue and Gall (11) with the addition of the high-speed scintillation method of Durie and Salmon (1). Kodak NTB-3 nuclear track emulsion was heated to 420C and then diluted with an equal weight of water, also at 42 C. The solution was allowed to sit without mixing (which creates air bubbles) for 10 min at 420C, then poured slowly into a Coplin jar (to facilitate slide dipping), and held at 420C. The slides with the hybridized [3H]cDNA were dipped slowly three times into the emulsion and air dried in a vertical position for 1 h. They were then dipped (10 s) in scintillator {5 g of PPO (2,5-diphenyloxazole), 100 mg of dimethyl PO- POP [1,4-bis-(5-phenyloxazolyl)-benzene] dissolved in 500 ml of dioxane}, air dried, sealed in light-proof black slide boxes, and exposed at -70 C for 3 weeks. After exposure, the slides were developed in Kodak D-19 developer at 17 C for 3 min, dipped in water for 10 s, dipped in Kodak fixer for 3 min, washed in water at 170C for 30 min, and air dried. The cells were stained with Wright-Giemsa (Harleco) for 8 min and for an additional 20 min after addition of ph 6.4 phosphate buffer, then rinsed with water, and air dried. It is essential to have adequate quantities of well-characterized cdna for this technique to be useful. For the synthesis of SiSV/SiSAV [3H]cDNA, 2 ml of 1,000x concentrated (1013 focus-forming units per ml) SiSV/SiSAV grown in marmoset cells (71AP1) was pelleted through a 30% glycerol (in 0.1 M NaCl-0.01 M Trishydrochloride, ph 7.4) J. VIROL. cushion at 100,000 x g for 1 h in a Beckman SW41 rotor. The virus pellet was resuspended in 500 yl of buffer: 2 mm dctp, 2 mm dttp, 0.6 mm [3H]dGTP, 0.9 mm [3H]dATP, 0.1% Triton X-100,4 mm magnesium acetate, 0.04 M Tris-hydrochloride (ph 7.4), 6 mm dithiothreitol, and 50 jig of actinomycin D per ml. After incubation on ice for 10 min, synthesis of [3H]cDNA was carried out at 370C for 16 h. The reaction was stopped by making the mixture 1% in sodium dodecyl sulfate and freezing at -70 C. After thawing, the mix was adjusted to ph 9, trna was added, and the cdna was extracted with an equal volume of 0.05 M Tris (ph 9)-saturated phenol-chloroform-isoamyl alcohol (1:1:0.01). After removal of the aqueous layer, the organic layer was repeatedly washed with 0.1 M NaCl-0.05% sodium dodecyl sulfate-0.02 M Tris (ph 9) until the interface collapsed. The combined aqueous layers were made 0.2 M with NaCl, 2 volumes of EtOH were added, and the cdna was precipitated overnight. The precipitate was collected by centrifugation, redissolved in 0.1 M NaCl-0.01 M Trischloride (ph 7.4) M EDTA, and nucleic acids were re-precipitated with cetyltrimethylammonium bromide (12). After a second EtOH precipitation, the cdna was alkali treated (0.3 N NaOH, 37 C overnight) and neutralized. Because of residual resistance (approximately 20%) to single-strand-specific S1 nuclease, the cdna was further purified by centrifugation on a neutral sucrose gradient (15 to 30% in 100 mm LiCl, 10 mm Tris (ph 7.4), 1 mm EDTA, and 0.2% sodium dodecyl sulfate) at 100,000 x g (30,000 rpm) for 16 h in a Beckman SW41 rotor. The main cdna peak (7S) was pooled, and the fractions at the top of the gradient were discarded. Approximately 4 x 107 cpm of cdna with a specific activity of 1.4 x 107 cpm/,ug was obtained from the synthesis. The cdna hybridized 75% to DNA from SiSV-infected cells at an uncorrected Cot of 8,500, with 6% self-annealing at the same Cot. When hybridized to SiSV ['25I]RNA,
3 VOL. 30, 1979 the cdna protected 100% of the RNA from RNase digestion at a cdna:rna ratio of 2:1, indicating that it was a representative copy of the entire RNA genome. Figure 1A shows the result of the hybridization of SiSV/SiSAV (71AP1) [3H]cDNA to 71AP1 cells producing SiSV/SiSAV. The grains, indicating the presence of hybridized [3H]cDNA, are dense over all the cells and almost negligible in intercellular spaces. The hybridization is specific: the addition of unlabeled competing SiSV/ SiSAV RNA (Fig. 1B), but not RNA from avian myeloblastosis virus, an unrelated virus (Fig. 1C), eliminates the grains. The hybridization is RNA dependent and is abolished when the cells are pretreated with RNase (Fig. 1D). Denaturation of cellular DNA (2) in 95% formamide- 0.1x SSC at 650C for 2 h before addition of [3H]cDNA resulted in no detectable hybridization, confirming that under the conditions used, htybridization was to cellular RNA. The specificity of the hybridization was also NOTE 639 demonstrated by the lack of hybridization of the SiSV/SiSAV (71AP1) [3H]cDNA to RNA from normal marmoset (HF) or normal human (A204) cell lines (Fig. 2A and 2B, respectively). There was also no hybridization seen between the SiSV/SiSAV (71AP1) [3H]cDNA and RNA of cells producing baboon endogenous virus (Fig. 2C) or cells producing feline leukemia virus (Fig. 2D). A bat cell line producing SiSV/SiSAV was as positive (Fig. 2E) as the 71AP1 (SiSV/SiSAV) cells used for production of the virus from which the cdna was synthesized, confirming that the positive hybridization was virus specific and not due to a marmoset cell-specific contaminant. Figure 3 shows the results of the hybridization of SiSV/SiSAV (71AP1) [3H]cDNA to 71AP1 (SiSV/SiSAV) cells diluted with uninfected feline embryo fibroblasts (FEF). The virus-producing cells are clearly detectable in the midst of the negative, nonproducing FEF cells. It is possible to detect one virus-positive cell on the slide, even if there are 103 or more nonproducing I FIG. 1. In situ hybridization of SiSV/SiSAV[3H]cDNA to the RNA of 71AP1 marmoset cells producing SiSV/SiSAV. Cytological preparations, [3H]cDNA synthesis, the hybridization conditions, and autoradiography were carried out as described in the text. Viral 70S RNA from SiSV/SiSA V and avian myeloblastosis virus was prepared as described in (16). (A) SiSV/SiSAV [3H]cDNA hybridized to 71AP1 cells producing SiSV/SiSAV; (B) same as (A) with the addition of competing, homologous SiSVISiSA V viral RNA; (C) same as (A) with the addition of competing, nonhomologous avian myeloblastosis virus RNA; (D) SiSV/SiSAV [3H]cDNA hybridized to 71AP1 (SiSV/SiSAV) cells which had been pretreated with RNase (1 IL of 1 mg of RNase A [Worthington]per ml and 2 [L of3x SSC were applied to the cells, covered, incubated in mineral oil at 37 C for 1 h, removed from oil, and washed as described in the text).
4 640 NOTE J. VIROL. ^S. A B V-.- :.. j....f. :. it v w C 1O.O.Aw. D Downloaded from FIG. 2. In situ hybridization of SiSV/SiSAV [3H]cDNA prepared from virus produced by marmoset cells to the RNA of cells not producing SiSVISiSA V or to nonmarmoset cells producing SiSV/SiSA V. (A) Normal marmoset cells (HF); (B) human rhabdomyosarcoma cells (A204); (C) A204 cellsproducing baboon endogenous virus (M); (D) FEF cells producing feline leukemia virus; (E) bat cells (B88) producing SiSV/SiSA V. cells also present. In situ hybridization of viral [3H]cDNA to cellular RNA should be useful for detecting cells replicating type C viruses in a mixed population or cells synthesizing viral RNA in a nonproducing system. The method is specific for viral RNA, has a low background that does not interfere with the detection of low levels of positive hybridization, and is independent of the number of virus-negative cells in the population. This makes it a very sensitive method to use in cell culture systems (viral induction, for example) where there is a small number of cells or where only a few cells might be expected to produce virus. Standard liquid hybridization systems might not be sensitive enough to detect such small amounts of viral RNA. We have also successfully hybridized SiSV/SiSAV [3H]cDNA to RNA from SiSV/SiSAV-infected cells which were grown directly on microscope slides (unpublished data). This technique has the advantage of making it possible to examine all the cells in a given population without the possibility of damage or loss of cells during cell washing and cytospin procedures. Perhaps the most interesting application of in on July 20, 2018 by guest
5 VOL. 30, '. *~~~~~~~~~~~. *-w.*. : %.,i.\..,%6 FIG. 3. In situ hybridization of SiSV/SiSAV [3H]cDNA to a mixture of virus-producing 71AP1 (SiSV/SiSAV9 cells and uninfected, nonvirus-producing FEF cells. Positive hybridization to the viruspositive cell is evident. situ hybridization in the RNA tumor virus system, however, is the examination of fresh cells from animals with virus-induced leukemia or lymphoma. It should be possible, by hybridizing viral cdna to tissue slices as well as to cytospins of peripheral blood and bone marrow samples, to identify tissues or cell populations which are sites of viral replication. The course of primary virus replication and subsequent spread could be followed during the course of such diseases as AKR leukemia and Friend erythro-leukemia in mice, feline leukemia virus-induced leukemia in cats, or natural and gibbon ape leukemia virus-induced leukemia in gibbon apes. It will hopefully be a useful tool in detecting cells which are expressing viral RNA in leukemia animals, or possibly in humans, when no replicating virus can be detected. Several SiSV-transformed nonproducer cell lines (obtained from C. Bergholz and S. Aaronson) are currently under examination to explore this possibility. The method also lends itself to the use of subfractionated cdna, enriched for specific regions of the viral genome, to investigate viral RNA expression in infected cells not producing virus. ACKNOWLEDGMENTS We thank Wolf Prensky and Paul Szabo of Sloan Kettering Institute and Linda Vaught and Brian Durie of the University of Arizona for information on the technical aspects of in situ hybridization and high-speed scintillation autoradiography, respectively; F. Ruscetti and M. Reitz for helpful discussion; R. Williams for excellent technical asistance; and A. Perry for infornation about the production of high titer SiSV/SiSAV (71AP1) virus #jb.,'pt.*.c..-.;r A -Z.,.:. I is NOTE 641 LITERATURE CITED 1. Durie, B. G. M., and S. E. Salmon High speed scintillation autoradiography. Science 190: Haase, A. T., L. Stowring, 0. Narayan, D. Griffin, and D. Price Slow persistent infection caused by visna virus: role of host restriction. Science 195: Harrison, P. R., D. Conkie, J. Paul, and K. Jones Localization of cellular globin messenger RNA by in situ hybridization to complementary DNA. FEBS Lett. 32: Hennig, W Molecular hybridization of DNA and RNA in situ. Int. Rev. Cytol. 36: Jones, K. W The method of in situ hybridization, p In R. H. Pain and B. J. Smith (ed.), New techniques in biophysics and cell biology. J. Wiley & Sons, London. 6. Loni, M. C., and M. Green Detection of viral DNA sequences in adenovirus-transformed cells by in situ hybridization. J. Virol. 12: Loni, M. C., and M. Green Virus-specific DNA sequences in mouse and rat cells transformed by the harvey and moloney murine sarcoma viruses detected by in situ hybridization. Virology 63: McDougall, J. K., and D. A. Galloway Detection of viral nucleic acid sequences using in situ cytological hybridization. 7th Annual ICN-UCLA Symposia. Molecular and Cellular Biology. J. Supramol. Struct. (Suppl. 2), p a.McDougall, J. K., D. A. Galloway, and C. M. Fenoglio In situ cytological hybridization to detect herpes simplex virus RNA in human tissues, p In P. Chandra (ed.), Antiviral mechanisms and the control of neoplasia. Plenum Press, New York. 9. Moar, M. H., and K. W. Jones Detection of virusspecific DNA and RNA base-sequences in individual cells transformed or infected by adenovirus type 2. Int. J. Cancer 16: Neer, A., N. Bara, and H. Manor In situ hybridization analysis of polyoma DNA replication in an inducible line of polyoma-transformed cells. Cell 11: Pardue, M. L., and J. G. Gall Nucleic acid hybridization to the DNA of cytological preparations. Methods Cell Biol. 11: Reitz, M. S., J. Abrell, C. D. Trainor, and R. C. GaBo Precipitation of nucleic acids with cetyltrimethylammonium bromide: a method for preparing viral and cellular DNA polymerase products for cesium sulfate density gradient analysis. Biochem. Biophys. Res. Commun. 49: Segal, S., M. Garner, M. F. Singer, and M. Rosenberg In situ hybridization of repetitive monkey genome sequences isolated from defective simian virus 40 DNA. Cell 9: Szabo, P., R. Elder, D. M. Steffensen, and 0. C. Uhlenbeck Quantitative in situ hybridization of ribosomal RNA species to polytene chromosomes of Drosophila melanogaster. J. Mol. Biol. 115: Wimber, D. E., and D. M. Steffensen Localization of gene function. Annu. Rev. Genet. 7: Wu, A., M. S. Reitz, M. Paran, and R. C. Gallo Mechanism of stimulation of murine type-c RNA tumor virus production by glucocorticoids post-transcriptional effects. J. Virol. 14:
Whole Mount Drosophila Embryo In Situ Hybridization with RNA probes 2/5/2001 Leslie Vosshall
Whole Mount Drosophila Embryo In Situ Hybridization with RNA probes 2/5/2001 Leslie Vosshall DAY ONE All incubations are done at room temperature unless otherwise noted. All solutions and all containers
More informationE.Z.N.A. SQ Blood DNA Kit II. Table of Contents
E.Z.N.A. SQ Blood DNA Kit II Table of Contents Introduction and Overview...2 Kit Contents/Storage and Stability...3 Blood Storage and DNA Yield...4 Preparing Reagents...5 100-500 μl Whole Blood Protocol...6
More informationGibbon Ape Leukemia Virus-Hall's Island: New Strain of Gibbon Ape Leukemia Virus
JOURNAL OF VIROLOGY, Jan. 1979, p. 395-400 0022-538X/79/0001-0395/06$02.00/0 Vol. 29, No. 1 Gibbon Ape Leukemia Virus-Hall's Island: New Strain of Gibbon Ape Leukemia Virus MARVIN S. REITZ, JR.,' FLOSSIE
More informationLoss of Proviral DNA Sequences in a Revertant of Kirsten Sarcoma Virus-transformed Murine Fibroblasts
J. gen. Virol. (I979), 44, 245-249 245 Printed in Great Britain Loss of Proviral DNA Sequences in a Revertant of Kirsten Sarcoma Virus-transformed Murine Fibroblasts (Accepted 22 February I979) SUMMARY
More informationSuperinfection with Vaccinia Virus
JOURNAL OF VIROLOGY, Aug. 1975, p. 322-329 Copyright 1975 American Society for Microbiology Vol. 16, No. 2 Printed in U.S.A. Abortive Infection of a Rabbit Cornea Cell Line by Vesicular Stomatitis Virus:
More informationProduct Manual. Omni-Array Sense Strand mrna Amplification Kit, 2 ng to 100 ng Version Catalog No.: Reactions
Genetic Tools and Reagents Universal mrna amplification, sense strand amplification, antisense amplification, cdna synthesis, micro arrays, gene expression, human, mouse, rat, guinea pig, cloning Omni-Array
More informationTranscription of the Marek's Disease Virus Genome in Virus-
JOURNAL OF VIROLOGY, Apr. 1979, p. 84-89 0022-538X/79/04-0084/06$02.00/0 Vol. 30, No. 1 Transcription of the Marek's Disease Virus Genome in Virus- Induced Tumors SANDRA SILVER, MARY SMITH, AND MEIHAN
More informationQuantitative Assay of Paravaccinia Virus Based
APPrU MICROBIOLOGY, JUly 1972, p. 138-142 Copyright 1972 American Society for Microbiology Vol. 24, No. 1 Printed in U.S.A. Quantitative Assay of Paravaccinia Virus Based on Enumeration of Inclusion-Containing
More informationvirus-i (RAV-1) or Rous associated virus-2 (RAV-2), do not transform but do produce
ISOLATION OF NONINFECTIOUS PARTICLES CONTAINING ROUS SARCOMA VIRUS RNA FROM THE MEDIUM OF ROUS SARCOMA VIRUS-TRANSFORMED NONPRODUCER CELLS* BY HARRIET LATHAM ROBINSONt VIRUS LABORATORY, UNIVERSITY OF CALIFORNIA,
More informationFormation of an Infectious Virus-Antibody Complex with Rous
JOURNAL OF VIROLOGY, Mar. 1976, p. 163-167 Copyright 1976 American Society for Microbiology Vol. 17, No. 3 Printed in U.S.A. Formation of an Infectious Virus-Antibody Complex with Rous Sarcoma Virus and
More informationChromatin IP (Isw2) Fix soln: 11% formaldehyde, 0.1 M NaCl, 1 mm EDTA, 50 mm Hepes-KOH ph 7.6. Freshly prepared. Do not store in glass bottles.
Chromatin IP (Isw2) 7/01 Toshi last update: 06/15 Reagents Fix soln: 11% formaldehyde, 0.1 M NaCl, 1 mm EDTA, 50 mm Hepes-KOH ph 7.6. Freshly prepared. Do not store in glass bottles. 2.5 M glycine. TBS:
More information10 mm KCl in a Ti-15 zonal rotor at 35,000 rpm for 16 hr at
Proc. Nat. Acad. SCi. USA Vol. 68, No. 11, pp. 2752-2756, November 1971 Translation of Exogenous Messenger RNA for Hemoglobin on Reticulocyte and Liver Ribosomes (initiation factors/9s RNA/liver factors/reticulocyte
More informationSize of Virus-Specific RNA in B-34, a Hamster Tumor Cell Producing Nucleic Acids of Type C Viruses from Three Species
JOURNAL OF VIROLOGY, OCt. 1975, p. 832-837 Copyright i 1975 American Society for Microbiology Vol. 16, No. 4 Printed in U.S.A. Size of Virus-Specific RNA in B-34, a Hamster Tumor Cell Producing Nucleic
More informationPRODUCT: RNAzol BD for Blood May 2014 Catalog No: RB 192 Storage: Store at room temperature
PRODUCT: RNAzol BD for Blood May 2014 Catalog No: RB 192 Storage: Store at room temperature PRODUCT DESCRIPTION. RNAzol BD is a reagent for isolation of total RNA from whole blood, plasma or serum of human
More informationChromatin Immunoprecipitation (ChIPs) Protocol (Mirmira Lab)
Chromatin Immunoprecipitation (ChIPs) Protocol (Mirmira Lab) Updated 12/3/02 Reagents: ChIP sonication Buffer (1% Triton X-100, 0.1% Deoxycholate, 50 mm Tris 8.1, 150 mm NaCl, 5 mm EDTA): 10 ml 10 % Triton
More informationHIV-1 Virus-like Particle Budding Assay Nathan H Vande Burgt, Luis J Cocka * and Paul Bates
HIV-1 Virus-like Particle Budding Assay Nathan H Vande Burgt, Luis J Cocka * and Paul Bates Department of Microbiology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, USA
More informationVIRUSES AND CANCER Michael Lea
VIRUSES AND CANCER 2010 Michael Lea VIRAL ONCOLOGY - LECTURE OUTLINE 1. Historical Review 2. Viruses Associated with Cancer 3. RNA Tumor Viruses 4. DNA Tumor Viruses HISTORICAL REVIEW Historical Review
More informationWilmington, Delaware cells were harvested in the cold and pelleted. The cell. pellet was suspended in 2 ml of cold buffer consisting
JOURNAL OF VIROLOGY, June 1969, p. 599-64 Vol. 3, No. 6 Copyright 1969 American Society for Microbiology Printed in U.S.A. Sindbis Virus-induced Viral Ribonucleic Acid Polymerasel T. SREEVALSAN' AND FAY
More informationMouse primary keratinocytes preparation
Mouse primary keratinocytes preparation 1. Fill a 150 X 25 mm petri dish with ice. Put newborn mice (2 3 days old) in the petri dish and insert it in an ice bucket. Leave the mice in the ice bucket for
More informationSeparation of sarcoma virus-specific and leukemia virus-specific genetic sequences of Moloney sarcoma virus (mechanism of transformation)
Proc. Nat. Acad. Sd. USA Vol. 72, No. 11, pp. 4650-4654, November 1975 Microbiology Separation of sarcoma virus-specific and leukemia virus-specific genetic sequences of Moloney sarcoma virus (mechanism
More informationSingle Cell Quantitative Polymer Chain Reaction (sc-qpcr)
Single Cell Quantitative Polymer Chain Reaction (sc-qpcr) Analyzing gene expression profiles from a bulk population of cells provides an average profile which may obscure important biological differences
More informationReplication of Sindbis Virus V. Polyribosomes and mrna in Infected Cells
JOURNAL OF VIROLOGY, Sept. 1974, p. 552-559 Vol. 14, No. 3 Copyright @ 1974 American Society for Microbiology Printed in U.S.A. Replication of Sindbis Virus V. Polyribosomes and mrna in Infected Cells
More informationSearch for viral nucleic sequences in rheumatoid cells
Annals of the Rheumatic Diseases, 1979, 38, 456-462 Search for viral nucleic sequences in rheumatoid cells MARY NORVAL1 AND CAROL SMITH2 From the 1Department of Bacteriology, University of Edinburgh Medical
More information(EDTA))." This preparation contained dsrna-1, dsrna-2, SPECIFICITY IN TRANSCRIPTION OF THE REOVIRUS GENOME*
SPECIFICITY IN TRANSCRIPTION OF THE REOVIRUS GENOME* BY Y. WATANABE, L. PREVECt AND A. F. GRAHAM THE WISTAR INSTITUTE OF ANATOMY AND BIOLOGY, PHILADELPHIA, PENNSYLVANIA Communicated by Thomas F. Anderson,
More informationSUPPLEMENTARY INFORMATION. Bacterial strains and growth conditions. Streptococcus pneumoniae strain R36A was
SUPPLEMENTARY INFORMATION Bacterial strains and growth conditions. Streptococcus pneumoniae strain R36A was grown in a casein-based semisynthetic medium (C+Y) supplemented with yeast extract (1 mg/ml of
More informationthe xenotropic sequences in the region of the env gene. The to the env gene of mouse xenotropic type C virus.
Proc. Natl. Acad. Sci. U$A Vol. 74, No. 10, pp. 4671-4675, October 1977 Microbiology Friend strain of spleen focus-forming virus is a recombinant between ecotropic murine type C virus and the env gene
More informationFOCUS SubCell. For the Enrichment of Subcellular Fractions. (Cat. # ) think proteins! think G-Biosciences
169PR 01 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name FOCUS SubCell For the Enrichment of Subcellular Fractions (Cat. # 786 260) think
More informationTrident Membrane Protein Extraction Kit
Cat. No. Size Shelf life GTX16373 5/ 20 tests 12 months at the appropriate storage temperatures (see below) Contents Component Storage Amount for 5 tests Amount for 20 tests Buffer A -20 o C 2.5 ml 10
More informationMidi Plant Genomic DNA Purification Kit
Midi Plant Genomic DNA Purification Kit Cat #:DP022MD/ DP022MD-50 Size:10/50 reactions Store at RT For research use only 1 Description: The Midi Plant Genomic DNA Purification Kit provides a rapid, simple
More informationAdjust to ph=7 with some HCl if too basic, or NaOH if too acid (but it will be quite close to neutrality even without adding any of these)
BIG VOLUME SOLUTIONS Solutions should be prepared with distilled and autoclaved water unless otherwise indicated. If the solutions have to be left unused for long periods (> 1 week), it is recommended
More informationFayth K. Yoshimura, Ph.D. September 7, of 7 RETROVIRUSES. 2. HTLV-II causes hairy T-cell leukemia
1 of 7 I. Diseases Caused by Retroviruses RETROVIRUSES A. Human retroviruses that cause cancers 1. HTLV-I causes adult T-cell leukemia and tropical spastic paraparesis 2. HTLV-II causes hairy T-cell leukemia
More informationAnnexure III SOLUTIONS AND REAGENTS
Annexure III SOLUTIONS AND REAGENTS A. STOCK SOLUTIONS FOR DNA ISOLATION 0.5M Ethylene-diamine tetra acetic acid (EDTA) (ph=8.0) 1M Tris-Cl (ph=8.0) 5M NaCl solution Red cell lysis buffer (10X) White cell
More informationEncapsidation of Sendai Virus Genome RNAs by Purified
JOURNAL OF VIROLOGY, Mar. 1988, p. 834-838 22-538X/88/3834-5$2./ Copyright C) 1988, American Society for Microbiology Vol. 62, No. 3 Encapsidation of Sendai Virus Genome RNAs by Purified NP Protein during
More informationWork-flow: protein sample preparation Precipitation methods Removal of interfering substances Specific examples:
Dr. Sanjeeva Srivastava IIT Bombay Work-flow: protein sample preparation Precipitation methods Removal of interfering substances Specific examples: Sample preparation for serum proteome analysis Sample
More informationAntigenic Analysis of Isolated Polypeptides from Visna Virus
INFECTION AND IMMUNITY, June 1976, p. 1728-1732 Copyright 1976 American Society for Microbiology Vol. 13, No. 6 Printed in USA. Antigenic Analysis of Isolated Polypeptides from Visna Virus P. D. MEHTA,*
More informationInhibition of reverse transcriptases by seminalplasmin
Biochem. J. (1983) 29, 183-188 183 Printed in Great Britain Inhibition of reverse transcriptases by seminalplasmin E. Shyam Prasad REDDY,* M. Ramachandra DAS,* E. Premkumar REDDYt and Pushpa M. BHARGAVA*
More informationMasterPure RNA Purification Kit
Cat. No. MCR85102 The MasterPure RNA Purification Kit pro vides all of the reagents necessary to recover RNA from a wide variety of biological sources. This kit uses a rapid desalting process 1 to remove
More informationFor purification of viral DNA and RNA from a wide range of sample materials
QIAamp virus kits For purification of viral DNA and RNA from a wide range of sample materials Automatable on QIAGEN s proven QIAamp Kits set the standard for purification of viral DNA and RNA. QIAamp virus
More informationATTACHMENT OF RIBOSOMES TO MEMBRANES DURING POLYSOME FORMATION IN MOUSE SARCOMA 180 CELLS
Published Online: 1 June, 1971 Supp Info: http://doi.org/10.1083/jcb.49.3.683 Downloaded from jcb.rupress.org on November 2, 2018 ATTACHMENT OF RIBOSOMES TO MEMBRANES DURING POLYSOME FORMATION IN MOUSE
More informationBiochemical Studies on Adenovirus Multiplication
JOURNAL OF VIROLOGY, OCt. 1969, p. 423-428 Copyright 1969 American Society for Microbiology Vol. 4, No. 4 Prinited ilt U.S.A. Biochemical Studies on Adenovirus Multiplication XVI. Transcription of the
More informationsive NIH/3T3 or BALB/8T3 cells by the XC plaque assay (9). Virus Infection and Infective Center Assay. Subconfluent
Proc. Nati. Acad. Sci. USA Vol. 73, No. 7, pp. 2236-224, July 1976 Biochemistry Effect of Fv-1 gene product on proviral DNA formation and integration in cells infected with murine leukemia viruses (N-tropic
More informationTRANSPORT OF AMINO ACIDS IN INTACT 3T3 AND SV3T3 CELLS. Binding Activity for Leucine in Membrane Preparations of Ehrlich Ascites Tumor Cells
Journal of Supramolecular Structure 4:441 (401)-447 (407) (1976) TRANSPORT OF AMINO ACIDS IN INTACT 3T3 AND SV3T3 CELLS. Binding Activity for Leucine in Membrane Preparations of Ehrlich Ascites Tumor Cells
More informationIdentification of the Virucidal Agent in Wastewater Sludge
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Apr. 1977, p. 860-864 Copyright X) 1977 American Society for Microbiology Vol. 33, No. 4 Printed in U.S.A. Identification of the Virucidal Agent in Wastewater Sludge
More informationVirus Harvest AAV 15 cm 2
Virus Harvest AAV 15 cm 2 1) Gently scrape cells from plate in 10 ml of media, place remaining 10 ml of media in plate lid. 2) Once cells are removed from plate, wash plate by pipetting 20 ml of media
More information[4 X 108 plaque-forming units (PFU)/ml] except in the experiment
Proc. Nat. Acad. Sci. USA Vol. 69, No. 9, pp. 2404-2409, September 1972 Extensive Symmetrical Transcription of Simian Virus 40 DNA in Virus-Yielding Cells (SV40/monkey cells/actinomycin D/RNase/hybridization)
More informationPREPARATION OF IF- ENRICHED CYTOSKELETAL PROTEINS
TMM,5-2011 PREPARATION OF IF- ENRICHED CYTOSKELETAL PROTEINS Ice-cold means cooled in ice water. In order to prevent proteolysis, make sure to perform all steps on ice. Pre-cool glass homogenizers, buffers
More informationSequences in the 5 and 3 R Elements of Human Immunodeficiency Virus Type 1 Critical for Efficient Reverse Transcription
JOURNAL OF VIROLOGY, Sept. 2000, p. 8324 8334 Vol. 74, No. 18 0022-538X/00/$04.00 0 Copyright 2000, American Society for Microbiology. All Rights Reserved. Sequences in the 5 and 3 R Elements of Human
More informationMammalian Membrane Protein Extraction Kit
Mammalian Membrane Protein Extraction Kit Catalog number: AR0155 Boster s Mammalian Membrane Protein Extraction Kit is a simple, rapid and reproducible method to prepare cellular protein fractions highly
More informationSite on the RNA of an Avian Sarcoma Virus at Which Primer Is Bound
JOURNAL OF VIROLOGY, Sept. 1975, p. 553-558 Copyright 0 1975 American Society for Microbiology Vol. 16, No. 3 Printed in U.SA. Site on the RNA of an Avian Sarcoma Virus at Which Primer Is Bound JOHN M.
More informationPRODUCT INFORMATION & MANUAL
PRODUCT INFORMATION & MANUAL 0.4 micron for Overall Exosome Isolation (Cell Media) NBP2-49826 For research use only. Not for diagnostic or therapeutic procedures. www.novusbio.com - P: 303.730.1950 - P:
More informationFor the 5 GATC-overhang two-oligo adaptors set up the following reactions in 96-well plate format:
Supplementary Protocol 1. Adaptor preparation: For the 5 GATC-overhang two-oligo adaptors set up the following reactions in 96-well plate format: Per reaction X96 10X NEBuffer 2 10 µl 10 µl x 96 5 -GATC
More informationGenomic Alterations Associated with Persistent Infections by Equine Infectious Anaemia Virus, a Retrovirus
J. gen. Virol. (1984), 65, 1395-1399. Printed in Great Britain 1395 Key words: EIA V/retrovirus persistence~antigenic variation/oligonucleotide mapping Genomic Alterations Associated with Persistent Infections
More information19 Viruses BIOLOGY. Outline. Structural Features and Characteristics. The Good the Bad and the Ugly. Structural Features and Characteristics
9 Viruses CAMPBELL BIOLOGY TENTH EDITION Reece Urry Cain Wasserman Minorsky Jackson Outline I. Viruses A. Structure of viruses B. Common Characteristics of Viruses C. Viral replication D. HIV Lecture Presentation
More informationNEUTRALIZATION OF REOVIRUS: THE GENE RESPONSIBLE FOR THE NEUTRALIZATION ANTIGEN* BY HOWARD L. WEINER~ AN~ BERNARD N. FIELDS
NEUTRALIZATION OF REOVIRUS: THE GENE RESPONSIBLE FOR THE NEUTRALIZATION ANTIGEN* BY HOWARD L. WEINER~ AN~ BERNARD N. FIELDS (From the Department of Microbiology and Molecular Genetics, Harvard Medical
More informationTrypsin Mass Spectrometry Grade
058PR-03 G-Biosciences 1-800-628-7730 1-314-991-6034 technical@gbiosciences.com A Geno Technology, Inc. (USA) brand name Trypsin Mass Spectrometry Grade A Chemically Modified, TPCK treated, Affinity Purified
More informationThe following protocol describes the isolation of nuclei from tissue. Item. Catalog No Manufacturer
SOP: Nuclei isolation from tissue and DNaseI treatment Date modified: 090923 Modified by: P. Sabo. (UW) The following protocol describes the isolation of nuclei from tissue. Ordering Information Item.
More informationASSAY OF SPHINGOMYELINASE ACTIVITY
ASSAY OF SPHINGOMYELINASE ACTIVITY Protocol for Protein Extraction Stock Solution 1. Leupeptin/hydrochloride (FW 463.0,
More informationAnalysis of small RNAs from Drosophila Schneider cells using the Small RNA assay on the Agilent 2100 bioanalyzer. Application Note
Analysis of small RNAs from Drosophila Schneider cells using the Small RNA assay on the Agilent 2100 bioanalyzer Application Note Odile Sismeiro, Jean-Yves Coppée, Christophe Antoniewski, and Hélène Thomassin
More informationMinute TM Plasma Membrane Protein Isolation and Cell Fractionation Kit User Manual (v5)
Minute TM Plasma Membrane Protein Isolation and Cell Fractionation Kit Catalog number: SM-005 Description Minute TM plasma membrane (PM) protein isolation kit is a novel and patented native PM protein
More informationHarvey Sarcoma Virus: A Second Murine Type C Sarcoma
JOURNAL OF VIROLOGY, June 1974. p. 1211-1219 Copyright 1974 American Society for Microhiology Vol. 1;3, No. 6 Printed in U.S.A. Harvey Sarcoma Virus: A Second Murine Type C Sarcoma Virus with Rat Genetic
More informationSpecificity of the DNA Product of RNA-Dependent DNA Polymerase
Proc. Nat. Acad. Sci. USA Vol. 70, No. 12, Part II, pp. 3923-3927, December 1973 Specificity of the DNA Product of RNA-Dependent DNA Polymerase in Type C Viruses: II. Quantitative Analysis* (DNA-RNA hybridization/murine
More information2) What is the difference between a non-enveloped virion and an enveloped virion? (4 pts)
Micro 260 SFCC Spring 2010 Name: All diagrams and drawings shall be hand drawn (do not photo-copied from a publication then cut and pasted into work sheet). Do not copy other student s answers. Para phase
More informationEpstein-Barr Virus: Stimulation By 5 '-Iododeoxy uridine or 5 '-Brom odeoxy uridine in Human Lymphoblastoid Cells F ro m a Rhabdom yosarcom a*
A n n a ls o f C l i n i c a l L a b o r a t o r y S c i e n c e, Vol. 3, No. 6 Copyright 1973, Institute for Clinical Science Epstein-Barr Virus: Stimulation By 5 '-Iododeoxy uridine or 5 '-Brom odeoxy
More informationOrganic and biochemical synthesis of monolignol biosynthetic pathway intermediates
Jie Liu 2012-2-8 Organic and biochemical synthesis of monolignol biosynthetic pathway intermediates 1. Organic synthesis of 5-hydroxyferulic acid Malonic acid 3, 4-Dihydroxy-5-methoxy-benzaldehyde 0.1
More informationVirus Particles in Interferon-Treated NIH 3T3 Cells Chronically Producing Moloney Murine Leukemia Virus
JOURNAL OF VIROLOGY, Feb. 1983, p. 489-495 0022-538X/83/020489-07$02.00/0 Copyright 1983, American Society for Microbiology Vol. 45, No. 2 Accumulation and Breakdown of RNA-Deficient Intracellular Virus
More informationViruses Tomasz Kordula, Ph.D.
Viruses Tomasz Kordula, Ph.D. Resources: Alberts et al., Molecular Biology of the Cell, pp. 295, 1330, 1431 1433; Lehninger CD Movie A0002201. Learning Objectives: 1. Understand parasitic life cycle of
More informationOverview: Chapter 19 Viruses: A Borrowed Life
Overview: Chapter 19 Viruses: A Borrowed Life Viruses called bacteriophages can infect and set in motion a genetic takeover of bacteria, such as Escherichia coli Viruses lead a kind of borrowed life between
More information7.012 Problem Set 6 Solutions
Name Section 7.012 Problem Set 6 Solutions Question 1 The viral family Orthomyxoviridae contains the influenza A, B and C viruses. These viruses have a (-)ss RNA genome surrounded by a capsid composed
More informationOctober 26, Lecture Readings. Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell
October 26, 2006 Vesicular Trafficking, Secretory Pathway, HIV Assembly and Exit from Cell 1. Secretory pathway a. Formation of coated vesicles b. SNAREs and vesicle targeting 2. Membrane fusion a. SNAREs
More informationSubeellular Distribution of Newly Synthesized Virus-Specific Polypeptides in Moloney Murine Leukemia Virus- Infected Cells
JOURNAL OF VIROLOGY, Jan. 1979, p. 385-389 0022-538X/79/01-0385/05$02.00/0 Vol. 29, No. 1 Subeellular Distribution of Newly Synthesized Virus-Specific Polypeptides in Moloney Murine Leukemia Virus- Infected
More informationViral-Related RNA in Hodgkins' Disease and Other Human Lymphomas
Proc. Nat. Acad. Soc. USA Vol. 69, No. 7, pp. 1727-1731, July 1972 Viral-Related RNA in Hodgkins' Disease and Other Human Lymphomas (DNA-RNA hybridization/rauscher murine leukemia virus/cancer/neoplasia/tumor)
More informationwas prepared from FPV-infected CEF (6) and fractionated by 5-20% sucrose in LETS buffer (100 mm LiCl/10 mm Tris-HCl,
Proc. Natl. Acad. Sci. USA Vol. 76, No. 8, pp. 379-3794, August 1979 Biochemistry The smallest genome RNA segment of influenza virus contains two genes that may overlap (fowl plague virus/cell-free translation/peptide
More informationVIRUSES. 1. Describe the structure of a virus by completing the following chart.
AP BIOLOGY MOLECULAR GENETICS ACTIVITY #3 NAME DATE HOUR VIRUSES 1. Describe the structure of a virus by completing the following chart. Viral Part Description of Part 2. Some viruses have an envelope
More informationCocultivation as a Tool for the Detection of Oncovimses in Childhood Leukemia*
Cocultivation as a Tool for the Detection of Oncovimses in Childhood Leukemia* Nooter. K.l, Zurcher, C.2, Coolen, C.2, Bentvelzen, P.l 1 Radiobiological Institute TNO. 151 Lange Kleiweg. Rijswijk. The
More informationInfluenza A H1N1 HA ELISA Pair Set
Influenza A H1N1 HA ELISA Pair Set for H1N1 ( A/Puerto Rico/8/1934 ) HA Catalog Number : SEK11684 To achieve the best assay results, this manual must be read carefully before using this product and the
More informationMaterials and Methods , The two-hybrid principle.
The enzymatic activity of an unknown protein which cleaves the phosphodiester bond between the tyrosine residue of a viral protein and the 5 terminus of the picornavirus RNA Introduction Every day there
More informationItem Catalog Number Manufacturer 1,4-Dithioerythritol (1 g) D9680 Sigma-Aldrich
SOP: Nuclei isolation from fresh mouse tissues and DNaseI treatment Date modified: 01/12/2011 Modified by: E. Giste/ T. Canfield (UW) The following protocol describes the isolation of nuclei and subsequent
More informationViruses. Rotavirus (causes stomach flu) HIV virus
Viruses Rotavirus (causes stomach flu) HIV virus What is a virus? A virus is a microscopic, infectious agent that may infect any type of living cell. Viruses must infect living cells in order to make more
More informationRole of Interferon in the Propagation of MM Virus in L Cells
APPLIED MICROBIOLOGY, Oct. 1969, p. 584-588 Copyright ( 1969 American Society for Microbiology Vol. 18, No. 4 Printed in U S A. Role of Interferon in the Propagation of MM Virus in L Cells DAVID J. GIRON
More informationHepatitis B Virus Genemer
Product Manual Hepatitis B Virus Genemer Primer Pair for amplification of HBV Viral Specific Fragment Catalog No.: 60-2007-10 Store at 20 o C For research use only. Not for use in diagnostic procedures
More informationEach Other. EDTA), quickly cooled in an ice slurry, and made 3 M KCl. before being bound to the column. Sindbis virus RNAs (49S
JOURNAL OF VIROLOGY, Mar. 1986, p. 917-921 0022-538X/86/030917-05$02.00/0 Vol. 57, No. 3 RNA Virus Genomes Hybridize to Cellular rrnas and to Each Other MARCELLA A. McCLURElt* AND JACQUES PERRAULT'2: Department
More informationInfectious Process of the Parvovirus H-1: Correlation of Protein Content, Particle Density, and Viral Infectivity
JOURNAL OF VIROLOGY, Sept. 1981, P. 800-807 Vol. 39, No. 3 0022-538X/81/090800-08$02.00/0 Infectious Process of the Parvovirus H-1: Correlation of Protein Content, Particle Density, and Viral Infectivity
More informationTranscription and RNA processing
Transcription and RNA processing Lecture 7 Biology 3310/4310 Virology Spring 2018 It is possible that Nature invented DNA for the purpose of achieving regulation at the transcriptional rather than at the
More informationManual (Second edition)
Reagent for RNA Extraction ISOGENⅡ Manual (Second edition) Code No. 311-07361 Code No. 317-07363 NIPPON GENE CO., LTD. Table of contents I Product description 1 II Product content 1 III Storage 1 IV Precautions
More informationTransfection of Sf9 cells with recombinant Bacmid DNA
Transposition Bacmid DNA Mini Culturing baculo cells Transfection of Sf9 cells with recombinant Bacmid DNA Amplification of the virus Titration of baculo stocks Testing the expression Transposition 1.
More informationSupplementary material: Materials and suppliers
Supplementary material: Materials and suppliers Electrophoresis consumables including tris-glycine, acrylamide, SDS buffer and Coomassie Brilliant Blue G-2 dye (CBB) were purchased from Ameresco (Solon,
More informationSputum DNA Collection, Preservation and Isolation Kit 50 Individual Devices
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Sputum DNA Collection, Preservation and Isolation Kit 50 Individual
More informationPinpoint Slide RNA Isolation System II Catalog No. R1007
INSTRUCTION MANUAL Pinpoint Slide RNA Isolation System II Catalog No. R1007 Highlights Allows for the isolation of total RNA from paraffin-embedded tissue sections on glass slides Simple procedure combines
More informationNorgen s HIV proviral DNA PCR Kit was developed and validated to be used with the following PCR instruments: Qiagen Rotor-Gene Q BioRad icycler
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com HIV Proviral DNA PCR Kit Product # 33840 Product Insert Background Information
More informationThawing MEFs (Mouse Embryonic Fibroblasts (MEFs)
1 FEEDER CULTURES The function of feeder cultures is to support the undifferentiated growth of hpsc. Typically primary fibroblasts are used for this purpose. We prepare our mouse feeder cells from ICR
More informationSTUDIES OF THE HEMAGGLUTININ OF HAEMOPHILUS PERTUSSIS HIDEO FUKUMI, HISASHI SHIMAZAKI, SADAO KOBAYASHI AND TATSUJI UCHIDA
STUDIES OF THE HEMAGGLUTININ OF HAEMOPHILUS PERTUSSIS HIDEO FUKUMI, HISASHI SHIMAZAKI, SADAO KOBAYASHI AND TATSUJI UCHIDA The National Institute of Health, Tokyo, Japan (Received: August 3rd, 1953) INTRODUCTION
More informationPhosphate buffered saline (PBS) for washing the cells TE buffer (nuclease-free) ph 7.5 for use with the PrimePCR Reverse Transcription Control Assay
Catalog # Description 172-5080 SingleShot Cell Lysis Kit, 100 x 50 µl reactions 172-5081 SingleShot Cell Lysis Kit, 500 x 50 µl reactions For research purposes only. Introduction The SingleShot Cell Lysis
More informationEVALUATION OF THE EFFECTIVENESS OF A 7% ACCELERATED HYDROGEN PEROXIDE-BASED FORMULATION AGAINST CANINE PARVOVIRUS
Final report submitted to Virox Technologies, Inc. EVALUATION OF THE EFFECTIVENESS OF A 7% ACCELERATED HYDROGEN PEROXIDE-BASED FORMULATION AGAINST CANINE PARVOVIRUS Syed A. Sattar, M.Sc., Dip. Bact., M.S.,
More informationDeterminants of the Host Range of Feline Leukaemia Viruses
J. gen. Virol. (1973), 20, I69-t75 Printed in Great Britain 169 Determinants of the Host Range of Feline Leukaemia Viruses By O. JARRETT, HELEN M. LAIRD AND D. HAY University of Glasgow, Leukaemia Research
More informationMEK1 Assay Kit 1 Catalog # Lot # 16875
MEK1 Assay Kit 1 Kit Components Assay Dilution Buffer (ADB), Catalog # 20-108. Three vials, each containing 1.0ml of assay dilution buffer (20mM MOPS, ph 7.2, 25mM ß-glycerol phosphate, 5mM EGTA, 1mM sodium
More informationMethodology for the Extraction of Brain Tissue Protein. Learning Objectives:
Proteomics Extraction of Brain Tissue Protein Methodology for the Extraction of Brain Tissue Protein Extraction of the entire protein from the sample requires optimized protocol and many protocols have
More informationIntroduction.-Cytopathogenic viruses may lose their cell-destroying capacity
AN INHIBITOR OF VIRAL ACTIVITY APPEARING IN INFECTED CELL CULTURES* BY MONTO Hot AND JOHN F. ENDERS RESEARCH DIVISION OF INFECTIOUS DISEASES, THE CHILDREN'S MEDICAL CENTER, AND THE DEPARTMENT OF BACTERIOLOGY
More informationPepsin Solution ready-to-use
SIE HABEN DIE VISION, WIR HABEN DIE SUBSTANZ. Pepsin Solution Single component Pepsin Solution: only one component refrigerator stable Pepsin is a commonly used digestive enzyme for immunohistochemical
More informationEPIGENTEK. EpiQuik Global Histone H3 Acetylation Assay Kit. Base Catalog # P-4008 PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE
EpiQuik Global Histone H3 Acetylation Assay Kit Base Catalog # PLEASE READ THIS ENTIRE USER GUIDE BEFORE USE The EpiQuik Global Histone H3 Acetylation Assay Kit is suitable for specifically measuring global
More information