Viral RNA / DNA Purification Guide Viral RNA / DNA purification products from MACHEREY-NAGEL MN guide for viral RNA / DNA purification Multiple solutions for many needs Single or high throughput Human or animal samples Highly sensitive viral nucleic acid recovery from various samples Superior reproducibility and quality for reliable downstream applications the best way to start your virus detection
MACHEREY-NAGEL viral RNA / DNA Kit Finder Mini NucleoSpin Virus / NucleoSpin Dx Virus p. 3 / 4 Cell-free fluid Blood, serum, plasma, tissue homogenate, feces, swab solution Silica membrane Magnetic beads Magnetic beads Funnel 8 / 96-well NucleoSpin RNA Virus F p. 4 NucleoSpin 8 / 96 Virus p. 5 NucleoMag 96 Virus p. 6 NucleoMag VET p. 7 Technologies NucleoSpin technology / Separation principle Procedure Features / Result Silica-membrane technology / Chaotropic salt binding Spin columns 8-well strips 96-well plates Low-throughput systems Medium-throughput systems High-throughput systems Principle: binding (high-salt) washing elution (low-salt) Adsorption of DNA / RNA in the presence of chaotropic salts (hydrate shell of DNA / RNA is reversibly removed) High-salt / ethanolic washing steps to remove contaminants Low-salt or water elution (hydrate shell is recovered, DNA / RNA is released from the membrane) Ready-to-use, sequencing and PCR-grade DNA / RNA From extra small to extra large scale, low to high throughput No alcohol precipitation necessary Fast and easy procedure NucleoSpin principle DNA / RNA contaminants DNA / RNA is bound to the silica membrane under high-salt conditions Contaminants are washed away under high-salt and / or ethanolic conditions DNA / RNA is eluted in low-salt buffer or water NucleoMag technology / Separation principle Procedure Features / Result Magnetic-bead technology / Chaotropic salt binding Flexible Principle: binding (high-salt) washing elution (low-salt) Adsorption of DNA / RNA in the presence of chaotropic salts (hydrate shell of DNA / RNA is reversibly removed) High-salt / ethanolic washing steps to remove contaminants Low-salt or water elution (hydrate shell is recovered, DNA / RNA is released from the beads) Highly-pure ready-to-use PCR products, genomic DNA / RNA Easily adaptable for automated use, e.g., on KingFisher, KingFisher ml, KingFisher 96, and KingFisher Flex NucleoMag principle DNA / RNA contaminants NucleoMag Beads NucleoMag Beads and binding buffer are added to the sample DNA / RNA is bound to the NucleoMag Beads Beads are held in the well by the magnet while contaminants are washed away DNA / RNA is eluted from the beads and recovered, while beads are held in the well by the magnet 2
NucleoSpin Virus Reliable purification of viral RNA and DNA from cell-free biological fluids XXOne kit for viral RNA and DNA Highly sensitive recovery of viral nucleic acid for reliable downstream application XXCompatible with liquid cell-free body fluids, such as serum or plasma Efficient recovery from 200 μl or up to 400 μl starting volume (two loading steps) Nucleic acid isolation from human and animal samples XXEfficient sample lysis Liquid Proteinase K is included Fragment size Silica-membrane technology Mini spin columns 200 μl serum, plasma, cell-free biological fluids (up to 400 μl with two loading steps) 100 bp approx. 50 kb 30 μl Application data 35,00 33,00 37 36 R MN 31,00 29,00 35 C T value 27,00 25,00 27,16 27,55 C T value 34 33 23,00 32 21,00 19,00 MN High sensitive viral BTV RNA detection with NucleoSpin Virus kit Viral BTV RNA was isolated from 12 different bovine EDTA-blood samples with different virus titer using NucleoSpin Virus and a competitor kit Q. BTV RNA was quantified by Real-time RT-PCR. The average C T value is lower for NucleoSpin Virus indicating higher viral RNA yields. Data kindly provided by Dr. Hoffmann, Friedrich-Loeffler-Institut, Germany Q 31 30 100 1000 10000 Input CMV [IU/mL] Reliable viral CMV DNA recovery with NucleoSpin Virus kit NucleoSpin Virus and a competitor kit R were used for viral CMV DNA isolation from different plasma samples with diverse virus titer. The viral DNA were quantified by qpcr in Roche LightCycler 480. The NucleoSpin Virus shows consistent reliable CMV DNA recovery. Data kindly provided by Dr. Tiemann, Dipl. Biol. Hartmann, LABCON-OWL GmbH, Germany NucleoSpin Virus* 10 / 50 / 250 740983.10 /.50 /.250 * The former NucleoSpin RNA Virus kit is still available (REF 740956.10 /.50 /.250). 3
NucleoSpin RNA Virus F Viral RNA and DNA* from biological fluids XXFor sample volumes up to 1 ml Patented technology of NucleoSpin Funnel Columns - small elution volume of 50 100 μl XXReliable and reproducible isolation of viral RNA and DNA* Safe and established procedure XXCarrier RNA included for highest sensitivity in downstream application XXClosed system preventing cross-contamination for highest safety Fragment size Binding capacity Silica-membrane technology Funnel columns < 1 ml cell-free biological fluids (e.g., serum, plasma) 100 bp approx. 50 kbp 50 100 μl 30 μg NucleoSpin RNA Virus F 25 740958 * For parallel isolation of viral RNA and DNA, use of Proteinase K is required (not included in the kit; see ordering information, p. 8). JJ NucleoSpin Dx Virus Viral RNA / DNA from human plasma and serum for in-vitro diagnostic purposes XXCE-IVD marked in accordance with EU Directive 98/79/EC Compliant with IVD directives in the EU XXFits into in-vitro diagnostic workflows CE-marked viral RNA and DNA isolation from plasma and serum Can be combined with common enzymatic amplification and detection of viral RNA or DNA XXCompatible with fresh or frozen samples treated with EDTA or citrate XXHighly consistent viral RNA and DNA isolation Reproducible isolations from different titers for reliable downstream applications XXSuitable for animal samples Applicable for non-ivd veterinary purpose Fragment size Binding capacity Silica-membrane technology Mini spin columns 150 μl plasma or serum (fresh, frozen, EDTA, or citrate treated) 100 bp approx. 50 kbp 50 μl 40 μg NucleoSpin Dx Virus* 50 740895.50 * CE-IVD marked kit: not availiable in all countries, please inquire. 4
NucleoSpin 8 / 96 Virus Isolation of viral RNA and DNA in 8-well strip and 96-well plate format XXTime-saving parallel isolation of viral RNA / DNA for serum, plasma, or biological fluids Proteinase K and Carrier RNA included XXReliable and reproducible isolation of viral RNA and DNA Free of impurities; MN Wash Plate minimizes risk of cross-contamination XXProcessing under vacuum or by centrifugation Suitable for manual and automated processing XXNucleoSpin 8 / 96 Virus Core Kit: Kits with basic content focused on automation platforms Additional accessories can be combined as needed NucleoSpin 8 Virus NucleoSpin 96 Virus Silica-membrane technology Silica-membrane technology 8-well strip 96-well plates < 150 μl cell-free biological fluids (e.g., serum, plasma) < 150 μl cell-free biological fluids (e.g., serum, plasma) Fragment size 100 bp approx. 50 kbp 100 bp approx. 50 kbp 70 100 μl 70 100 μl Binding capacity 40 μg 40 μg Application data Highly sensitive detection of viral RNA HCV RNA was purified from 96 plasma samples (100 μl). A dilution series was processed using NucleoSpin 96 Virus (12 samples each). Percentage of positive RT-PCR samples at low virus titers are shown (COBAS AMPLICOR, Roche Diagnostics). NucleoSpin 8 Virus 12 x 8 / 60 x 8 740643 /.5 NucleoSpin 8 Virus Core Kit 48 x 8 740451.4 NucleoSpin 96 Virus 2 x 96 / 4 x 96 740691.2 /.4 NucleoSpin 96 Virus Core Kit 4 x 96 740452.4 5
NucleoMag 96 Virus Magnetic-bead based isolation of viral RNA and DNA from serum or plasma XXReliable viral RNA / DNA purification for reproducible results New Buffer MVL for efficient lysis Proteinase K and Carrier RNA included XXOne-tube processing minimizes risk of cross-contamination XXYield does not depend on elution volume Small elution volumes > 50 μl XXEasily adapted to automated use Binding capacity Magnetic-bead based technology Highly reactive superparamagnetic beads < 200 μl cell-free biological fluids (e.g., serum, plasma) 50 100 μl 0.4 μg/μl beads Application data 1.000.000 100.000 HCV detection R 2 = 0.9977 1.000.000 100.000 HBV detection R 2 = 0.9962 HCV IU/mL 10.000 1.000 100 HBV IU/mL 10.000 1.000 100 10 10 1 1 10 100 1000 10.000 100.000 1.000.000 10 100 1000 10.000 100.000 1.000.000 Input HCV IU/mL Input HBV IU/mL Buffer MV1 Buffer MVL New Lysis Buffer MVL, higher sensitivity in viral DNA and RNA detection Detection of viral RNA and DNA in Real-time PCR (Artus RealArt HCV RNA / HBV DNA; Roche LightCycler 480), purified by NucleoMag 96 Virus using Buffer MVL and Buffer MV1. Nucleic acids were purified from virus samples of different titers and used for quantification. NucleoMag 96 Virus 1 x 96 / 4 x 96 744800.1 /.4 6
NucleoMag VET Magnetic-bead based isolation of viral RNA and DNA from veterinary samples XXAllround kit for veterinary diagnostics One kit for common veterinary samples XXHigh sensitivity even with low viral titers Reliable isolation from different viral titers XXRobust one-tube processing minimizes risk of cross-contamination XXEasily adapted to automated use Maximum amount of starting material Binding capacity Magnetic-bead based technology Highly reactive superparamagnetic beads < 200 μl whole blood, serum, plasma, < 10 30 mg tissue (e.g., ear notches), < 200 μl feces, < 200 μl swab wash solution < 200 μl liquid / homogenized sample 50 100 μl 0.4 μg/μl beads Application data A B Sensitive detection of viral RNA and DNA from animal samples A) For purifications, serum samples from PRRSV (blue) and PCV2 (red) positive animal samples were diluted 10-fold. Viral RNA (PRRSV) or viral DNA (PVC2) were isolated from 200 μl blood serum sample using NucleoMag VET kit. Purified RNA (PRRSV) was analyzed using Real-time Multiplex RT- PCR (VIROTYPE PRRSV, LDL). Purified DNA (PVC2) was analyzed using Real-time PCR (PCV2 Primer). Data was generated in cooperation IVD GmbH Hannover, Germany B) PCV2 positive animal tissue samples (n = 22, 20 mg each) were used for viral DNA purification using indicated kits. Purified viral DNA was analyzed by Real-time PCR (PCV2 Primer). Out of 22 samples 20 (MN) or 14 (Q) samples were tested positive with C T mean values shown in the graph. Data kindly provided by Italian customer NucleoMag VET 1 x 96 / 4 x 96 744200.1 /.4 7
Kits for purification of viral RNA and DNA Preps REF NucleoSpin Virus* 10 / 50 / 250 740983.10 /.50 /.250 NucleoSpin RNA Virus F 25 740958 NucleoSpin Dx Virus** 50 740895.50 NucleoSpin 8 Virus 12 x 8 / 60 x 8 740643 /.5 NucleoSpin 8 Virus Core Kit 48 x 8 740451.4 NucleoSpin 96 Virus 2 x 96 / 4 x 96 740691.2 /.4 NucleoSpin 96 Virus Core Kit 4 x 96 740452.4 NucleoMag 96 Virus 1 x 96 / 4 x 96 744800.1 /.4 NucleoMag VET 1 x 96 / 4 x 96 744200.1 /.4 Kits for purification of viral DNA from blood Preps REF NucleoSpin Blood 10 / 50 / 250 740951.10 /.50 /.250 Product accessories Pack of REF Liquid Proteinase K 5 ml 740396 Proteinase K 100 mg 740506 Carrier RNA 1 mg 740514.1000 Product accessories for NucleoSpin Pack of REF NucleoVac 96 Vacuum Manifold 1 740681 NucleoVac Vacuum Regulator 1 740641 Starter Set A (for use of NucleoSpin 8-well strips on NucleoVac 96 Vacuum Manifold) Starter Set C (for use of NucleoSpin 8-well strips under centrifugation) 1 740682 1 740684 Product accessories for NucleoMag Pack of REF NucleoMag SEP (magnetic separator; for use with 96-well plate) KingFisher 96 Accessory Kit A (Square-well Blocks, Deep-well Tip Combs, Elution Plates; for use with KingFisher 96 platform) 1 744900 1 set 744950 Material to be supplied by the user for NucleoMag Pack of REF Rack of Tube Strips (lysis tubes) 4 / 24 740477 /.24 Square-well Block 4 / 24 740481 /.24 Elution Plate U-bottom (elution plate) 24 740486.24 * The former NucleoSpin RNA Virus kit is still available (REF 740956.10 /.50 /.250). ** CE-IVD marked kit: not availiable in all countries, please inquire. Trademarks: MACHEREY-NAGEL: NucleoSpin and NucleoMag Roche: COBAS and LightCycler artus GmbH: RealArt QIAGEN Leipzig GmbH: VIROTYPE Thermo: KingFisher Your local distributor Image credits: spline_x Fotolia.com KATEN300150 / VirusGuide en1/5/0/06.2014 DD Printed in Germany MACHEREY-NAGEL EN ISO 9001: 2008 CERTIFIED MACHEREY-NAGEL GmbH & Co. KG Neumann-Neander-Str. 6 8 52355 Düren Germany Germany and international: Switzerland: MACHEREY-NAGEL AG France: MACHEREY-NAGEL EURL Tel.: +49 24 21 969-0 Tel.: +41 62 388 55 00 Tel.: +33 388 68 22 68 Fax: +49 24 21 969-199 Fax: +41 62 388 55 05 Fax: +33 388 51 76 88 E-mail: info@mn-net.com E-mail: sales-ch@mn-net.com E-mail: sales-fr@mn-net.com USA: MACHEREY-NAGEL Inc. Tel.: +1 484 821 0984 Fax: +1 484 821 1272 E-mail: sales-us@mn-net.com