DATA SHEET. Provided: 500 µl of 5.6 mm Tris HCl, 4.4 mm Tris base, 0.05% sodium azide 0.1 mm EDTA, 5 mg/liter calf thymus DNA.

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1 Viral Load

2 DNA >> Standard

3 PCR standard 0 Copies Catalog Number: 1122 Lot Number: Release Category: A Provided: 500 µl of 5.6 mm Tris HCl, 4.4 mm Tris base, 0.05% sodium azide 0.1 mm EDTA, 5 mg/liter calf thymus DNA. Description: 10 reactions of a negative control template for use alongside HIV-1 Z6 DNA PCR standards (Cat#'s ). -80 o C. Contributor: Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems. Produced by the VQA Laboratory at Rush University Medical Center. References: Hart C, Chant S-Y, Kwok S, Sninsky J, Ou C-Y, Schochetman G. A replication-deficient HIV-1 DNA used for quantitation of the polymerase chain reaction (PCR). Nucleic Acids Research 18: , Abstract Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, DAIDS, NIAID, NIH: PCR Standard 0 copies (cat# 1122) from Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems." Also include the reference cited above in any publications. REV: 12/10/2017 Page 1 of 1

4 PCR Standards 1 Copy Catalog Number: 4676 Lot Number: Release Category: A Provided: 500 µl of HIV-1 Z6 DNA at 1 copy/50 µl in 5.6 mm Tris HCl, 4.4 mm Tris base, 0.05% sodium azide 0.1 mm EDTA, 5 mg/liter calf thymus DNA. Description: 10 reactions of HIV-1 Z6 DNA at 1 copy/50 µl. -80 o C. Contributor: Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems. Produced by the VQA Laboratory at Rush University Medical Center. References: Hart C, Chant S-Y, Kwok S, Sninsky J, Ou C-Y, Schochetman G. A replication-deficient HIV-1 DNA used for quantitation of the polymerase chain reaction (PCR). Nucleic Acids Research 18: , Abstract Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, DAIDS, NIAID, NIH: PCR Standards 1 Copy (cat# 4676) from Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems." Also include the reference cited above in any publications. REV: 12/10/2017 Page 1 of 1

5 PCR Standards 20 Copies Catalog Number: 4677 Lot Number: Release Category: A Provided: 500 µl of HIV-1 Z6 DNA at 20 copies/50 µl in 5.6 mm Tris HCl, 4.4 mm Tris base, 0.05% sodium azide 0.1 mm EDTA, 5 mg/liter calf thymus DNA. Description: 10 reactions of HIV-1 Z6 DNA at 20 copies/50 µl. -80 o C. Contributor: Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems. Produced by the VQA Laboratory at Rush University Medical Center. References: Hart C, Chant S-Y, Kwok S, Sninsky J, Ou C-Y, Schochetman G. A replication-deficient HIV-1 DNA used for quantitation of the polymerase chain reaction (PCR). Nucleic Acids Research 18: , Abstract Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, DAIDS, NIAID, NIH: PCR Standards 20 Copies (cat# 4677) from Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems." Also include the reference cited above in any publications. REV: 12/10/2017 Page 1 of 1

6 PCR Standards 50 Copies Catalog Number: 4678 Lot Number: Release Category: A Provided: 500 µl of HIV-1 Z6 DNA at 50 copies/50 µl in 5.6 mm Tris HCl, 4.4 mm Tris base, 0.05% sodium azide 0.1 mm EDTA, 5 mg/liter calf thymus DNA. Description: 10 reactions of HIV-1 Z6 DNA at 50 copies/50 µl. -80 o C. Contributor: Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems. Produced by the VQA Laboratory at Rush University Medical Center. References: Hart C, Chant S-Y, Kwok S, Sninsky J, Ou C-Y, Schochetman G. A replication-deficient HIV-1 DNA used for quantitation of the polymerase chain reaction (PCR). Nucleic Acids Research 18: , Abstract Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, DAIDS, NIAID, NIH: PCR Standards 50 Copies (cat# 4678) from Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems." Also include the reference cited above in any publications. REV: 12/10/2017 Page 1 of 1

7 pvqa Catalog Number: Lot Number: Release Category: E Provided: 10µl at 10ng/µl in 10mM Tris. Description: A plasmid standard used for an HIV-1 mrna quantification assay. This assay utilizes a primer in the 3'LTR of HIV-1 and a poly-a anchored primer to specifically detect all poly-adenylated HIV-1 mrna species. The pvqa plasmid contains the last ~350 nucleotides of HIV-1 mrna as well as ~30 deoxyadenosine residues. Special Characteristics: This plasmid can be utilized as a DNA standard for qpcr reactions, or can be used as a template for in vitro transcription of an RNA standard for RT-qPCR. SEQUENCE -20 C Contributor: Dr. Robert Siliciano, Johns Hopkins School of Medicine References: Shan L, Rabi SA, Laird GM, Eisele EE, Zhang H, Margolick JB, Siliciano RF. A novel PCR assay for quantification of HIV-1 RNA. J Virol Jun;87(11): ABSTRACT Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: Cat# pvqa from Dr. Robert Siliciano." Also include the reference cited above in any publications. REV: 12/10/2017 Page 1 of 2

8 REV: 12/10/2017 Page 2 of 2

9 PCR Standards 100 Copies Catalog Number: 4679 Lot Number: Release Category: A Provided: 500 µl of HIV-1 Z6 DNA at 100 copies/50 µl in 5.6 mm Tris HCl, 4.4 mm Tris base, 0.05% sodium azide 0.1 mm EDTA, 5 mg/liter calf thymus DNA. Description: 10 reactions of HIV-1 Z6 DNA at 100 copies/50 µl. -80 C Contributor: Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems. Produced by the VQA Laboratory at Rush University Medical Center. References: Hart C, Chant S-Y, Kwok S, Sninsky J, Ou C-Y, Schochetman G. A replication-deficient HIV-1 DNA used for quantitation of the polymerase chain reaction (PCR). Nucleic Acids Research 18: , Abstract Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, DAIDS, NIAID, NIH: PCR Standards 100 Copies (cat# 4679) from Dr. Shirley Kwok and Dr. Cindy Christopherson, Roche Molecular Systems." Also include the reference cited above in any publications. REV: 12/10/2017 Page 1 of 1

10 HMA Kits >> Complete Kits

11 Heteroduplex Mobility Analysis HIV-1 gag Subtyping Kit Catalog Number: 5210 Lot Number: 9 Release Category: D Provided: The HMA Gag Kit includes primers, control plasmid DNA, and reference strains. Click here to view kit contents -70 C. Upon receipt, place the Primers and the Plasmids in separate boxes to avoid DNA cross contamination. Contributor: Drs. Leo Heyndrickx, Gert Van der Auwera, and Guido van der Groen. References: Website references: Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HMA HIV-1 Gag Subtyping Kit from Drs. Guido van der Groen, Leo Heyndrickx and Gert Van der Auwera." REV: 12/10/2017 Page 1 of 1

12 Heteroduplex Mobility Analysis HIV-1 env Subtyping Kit Catalog Number: 2751 Lot Number: Release Category: D Provided: The HMA Kit contains primers, control plasmid DNA, and reference strains. Click here to view kit contents -70 C. Upon receipt, place the Primers and the Plasmids in separate boxes to avoid DNA cross contamination. Contributor: Dr. James Mullins References: Website reference: Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: HMA HIV-1 Env Subtyping Kit from Dr. James Mullins." REV: 12/10/2017 Page 1 of 1

13 OLA Kits >> All

14 OLA Kit for the Detection of HIV-1 Resistance Mutations to NRTIs Catalog Number: 7645 Lot Number: Release Category: A Provided: Each kit contains plasmids, primers, codon-specific oligonucleotides and control genomic DNA. Click here to view kit contents Description: Used for the detection of resistance mutations to nucleoside reverse transcriptase inhibitors (abacavir, didanosine, lamivudine, stavudine, tenofovir, zalcitabine, zidovudine). OLA Kit Manual v C Contributor: Dr. Ingrid Beck, Dr. Lisa Frenkel, and DAIDS, NIAID. Acknowledgment for publications should read The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: OLA Kit, NRTIs (Cat #7645) from Dr. Ingrid Beck and Dr. Lisa Frenkel. REV: 12/10/2017 Page 1 of 1

15 OLA KIT for the Detection of HIV-1 Resistance Mutations to NNRTIs Catalog Number: 7644 Lot Number: Release Category: A Provided: Each kit contains plasmids, primers, codon-specific oligonucleotides and control genomic DNA. Click here to view kit contents Description: Used for the detection of HIV-1 resistance mutations to non-nucleoside reverse transcriptase inhibitors (nevirapine, delavirdine, efavirenz) OLA Kit Manual v C Contributor: Dr. Ingrid Beck, Dr. Lisa Frenkel, and DAIDS, NIAID. Acknowledgment for publications should read The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: OLA Kit, NNRTIs (Cat #7644) from Drs. Ingrid Beck and Lisa Frenkel. REV: 12/10/2017 Page 1 of 1

16 OLA Kit for the Detection of HIV-1 Resistance Mutations to Protease Inhibitors Catalog Number: 7643 Lot Number: 3 Release Category: A Provided: Each kit contains plasmid DNAs, primers, codon-specific oligonucleotides and control genomic DNA. Click here to view kit contents Description: Used for the detection of HIV-1 resistance mutations to protease inhibitors (nelfinavir, saquinavir, indinavir, ritonavir, and amprenavir). OLA Kit Manual v C Contributor: Drs. Ingrid Beck and Lisa Frenkel, and DAIDS, NIAID. Acknowledgment for publications should read The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: OLA Kit, Protease Inhibitors (Cat #7643), from Drs. Ingrid Beck and Lisa Frenkel. REV: 12/10/2017 Page 1 of 1

17 RNA >> Standard

18 AMΔHIV Catalog Number: 1913 Lot Number: 1 03/93 Release Category: A Provided: 1 ml cell-free virus Description: A 267 bp synthetic HIV-1MN gag sequence designed by Dr. Gregory Milman, DAIDS, NIAID, was cloned into the SalI-HindIII site of the retroviral vector pln6 by Drs. Janice Clements and Paula Pitha-Rowe (The Johns Hopkins University). Original Source: A 267 bp gag fragment (sequence supplied by Dr. Gregory Milman, DAIDS) was cloned into the synthetic HIV-1 MN SalI-HindIII sites of the retroviral vector plnl6. The viral particles were collected from infected PA317 cells transfected with the recombinant vector. Host: PA317 packaging cells. Titer: 2.0 x 10 7 copies/ml (Roche) 1.4 x 10 7 copies/ml (NASBA) Special Characteristics: This virus serves as an internal standard for detection of HIV RNA by RT-PCR. The gag fragment contains small deletions to distinguish it from wild type HIV-1 (see attached). SK38/SK39 or SK145/SK101 primers can be used for RT PCR. The amplified product can be distinguished from full-length HIV-1 gag RNA by liquid hybridizaion followed by gel electorphoresis. 1 µl of the virus is sufficient for PCR analysis and quantitation. Gag sequence Sterility: Negative for bacteria and mycoplasma. REV: 12/10/2017 Page 1 of 2

19 Liquid nitrogen. Contributor: Division of AIDS, NIAID, NIH, and Dr. Paula Pitha-Rowe. References: Miller AD, Buttimore C. Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol Cell Biol 6: , Bender MA, Palmer TD, Gelinas RE, Miller AD. Evidence that the packaging signal of Moloney murine leukemia virus extends into the gag region. J Virol 61: , Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, DAIDS, NIAID: AMΔHIV from Dr. Paula Pitha-Rowe." Also include the references cited above in any publications. REV: 12/10/2017 Page 2 of 2

20 HIV-1 VQA RNA Quantification Standard Catalog Number: 3443 Lot Number: Release Category: D Provided: 1.25 ml, 150,000 copies HIV-1 RNA/mL spiked into HIV negative human serum to obtain final concentration. Contains EDTA. Description: Quantified live infectious virus spiked into HIV negative human serum. Original Source: HIV-1 qualitative cultures from known positive donors. Special Characteristics: This virus concentrate can be used to quantify HIV-1 RNA for viral detection assays. Virus stock used for this standard is a pool of B subtype HIV cultured from the PBMCs of 5 separate HIV-infected donors who were naïve to antiretroviral treatment. The virus culture supernatants were clarified of all cellular material and pooled. The virus stock was diluted in a commercial pooled true normal serum that is also clarified by filtration and EDTA is added at a final concentration of 6mM to mirror negative human EDTA plasma. Liquid nitrogen. Contributor: DAIDS Virology Quality Assurance (VQA) Program, Rush Presbyterian-St. Luke s Medical Center, Chicago, IL. Director: Dr. James Bremer. References: Yen-Lieberman B, et al. Evaluation of a quality assurance program for quantitation of human immunodeficiency virus type 1 RNA in plasma by the AIDS Clinical Trials Group virology laboratories. J Clin Microbiol 34: , Abstract REV: 12/10/2017 Page 1 of 2

21 Acknowledgment for publications should read "The following reagent was obtained through the NIH AIDS Reagent Program, DAIDS, NIAID: HIV-1 VQA RNA Quantification Standard from the DAIDS Virology Quality Assurance (VQA) Program." Also include the reference cited above in any publications. REV: 12/10/2017 Page 2 of 2

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