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SV Total RNA Isolation System

SV Total RNA Isolation System

Extracts RNA from Tissues, Cultured Cells and White Blood Cells

  • Rapid isolation of high yields of total RNA without the use of hazardous compounds
  • Choice of microcentrifugation (spin) or vacuum purification methods
  • Purified RNA suitable for all routine molecular biology applications, including RT-PCR

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Description

Size

Catalog number: Z3101

Catalog number: Z3100

Catalog number:  Z3105

Isolate RNA with a Simple, 1-hour Procedure

The SV Total RNA Isolation System provides a fast and simple technique for preparation of intact total RNA from tissues, cultured cells and white blood cells in as little as one hour. Using this membrane-based purification system, up to 60mg of tissue can be processed per purification, depending on tissue type. The system incorporates a DNase treatment step directly on the minicolumn membrane. This step substantially reduces genomic DNA contamination, which can interfere with amplification-based methodologies. Purification is achieved without the use of phenol:chloroform extractions or ethanol precipitations, and there is no DNase carryover in the final RNA preparation

Sample Types Processed with the SV Total RNA Isolation System

Sample Type Promega External Comments
Fresh whole blood Yes Yes A protocol is provided in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Brisco, P. Sankbeil, J. and Kephart, D. (1997) Promega Notes 64, 7–10 Hara, N. et al. (2002) Clin. Cancer Res. 8, 1794–9. PubMed ID #12060619.
Human blood plasma and serum Yes Yes A protocol is provided in the SV Total RNA Isolation System Technical Manual #TM048. Additional resource: Fresh or once-frozen human serum (100μl) was added to 175μl of SV RNA Lysis Buffer, then the remainder of the SV Total RNA Isolation System protocol was followed. Chen, X.Q. et al. (2000) Clin. Cancer Res. 6, 3823–6. PubMed ID #11051224.
Human leukocytes Yes A protocol is provided in the SV Total RNA Isolation System Technical Manual #TM048.
Winter flounder red blood cells Yes Pedersen, S.F. et al. (2003) Am. J. Physiol. Cell Physiol. 284, C1561–76. PubMed ID #12734109.
Human peripheral blood mononuclear cells Yes Yawalker, N. et al. (1998) J. Invest. Dermatol. 111, 1053–7. PubMed ID #9856816. Escoffier, C. et al. (1999) J. Virol. 73, 1695–8. PubMed ID #9882382.
Primary CD4+ human T lymphocytes Yes Secchiero, P. et al. (2000) Proc. Natl. Acad. Sci. USA 97, 14620–5. PubMed ID #11114167.
Viral RNA from human plasma Yes Personal communication. Contact Promega Technical Services for more information.
Sample Type Promega External Comments
Mammalian tissues A general protocol for total RNA isolation from tissue is provided in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources are listed below
Mouse or rat brain Yes Yes Follow the standard protocol for tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Kephart, D. Measuring gene expression from mammalian brain tissue. eNotes Brisco, P. Sankbeil, J. and Kephart, D. (1997) Promega Notes 64, 7–10 Kephart, D. and Hooper, K. (1998) Promega Notes 69, 17–9. Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse thoracic dorsal root ganglion neurons Yes Ekström, P.A.R. et al. (2003) Mol. Cell. Neurosci. 22, 383–95. PubMed ID #12691739.
Mouse central nervous system Yes Serafini, B. et al. (2000) Am. J. Pathol. 157, 1991–2002. PubMed ID #11106572.
Mouse small intestine Yes Serafini, B. et al. (2000) Am. J. Pathol. 157, 1991–2002. PubMed ID #11106572. Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse muscle Yes Yes Follow the standard protocol for tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Sakai, H. et al. (1999) J. Physiol. 519, 323–33. PubMed ID #10457052. Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse or rat liver Yes Yes Follow the standard protocol for tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Sakai, H. et al. (1999) J. Physiol. 519, 323–33. PubMed ID #10457052. Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse kidney Yes Yes Follow the standard protocol for tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740. Attmane-Elakeb, A. et al. (2000) J. Biol. Chem. 275, 33548–53. PubMed ID #10942780.
Mouse spleen Yes Yes Follow the standard protocol for tissue in the SV Total RNA Isolation System Technical Manual #TM048. Serafini, B. et al. (2000) Am. J. Pathol. 157, 1991–2002. PubMed ID #11106572. Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse embryonic heart, brain and skeletal muscle Yes Agarkova, I. et al. (2000) J. Biol. Chem. 275, 10256–64. PubMed ID #10744711.
Mouse or rat lung Yes Yes Agarkova, I. et al. (2000) J. Biol. Chem. 275, 10256–64. PubMed ID #10744711.
Rat pancreas Yes Follow the standard protocol for tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Brisco, P. Sankbeil, J. and Kephart, D. (1997) Promega Notes 64, 7–10. Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse or rat heart Yes Yes Follow the standard protocol for tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Brisco, P. Sankbeil, J. and Kephart, D. (1997) Promega Notes 64, 7–10. Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse colon Yes Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse stomach Yes Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse thymus Yes Pinto, D. et al. (1999) J. Biol. Chem. 274, 6476–82. PubMed ID #10037740.
Mouse eyes Yes Lucas, R.J. et al. (1999) Science 284, 505–7. PubMed ID #10205062.
Mouse retina Yes Freedman, M.S. et al. (1999) Science 284, 502–4. PubMed ID #10205061.
Rat whole cochlea, modiola and cochlear sensorineural epithelial/lateral walls Yes Stöver, T. et al. (2000) Brain Res. Mol. Brain Res. 76, 25–35. PubMed ID #10719212.
Human muscle Yes Roth, S.M. et al. (2002) Physiol. Genomics 10, 181–90. PubMed ID #12209020.
Human placenta Yes Neuschäfer-Rube, F. et al. (2003) Biochem. J. 371, 443–9. PubMed ID #12519077. Gonzalez-Perrett, S. et al. (2001) Proc. Natl. Acad. Sci. USA 98, 1182–7. PubMed ID #11252306.
Human skin Yes Yawalker, N. et al. (1999) J. Invest. Dermatol. 113, 43–8. PubMed ID #10417617. Yawalker, N. et al. (1998) J. Invest. Dermatol. 111, 1053–7. PubMed ID #9856816.
Human brain Yes Miyasaka, T. et al. (2001) Am. J. Pathol. 158, 373–9. PubMed ID #11159174.
Primary human peritoneal mesothelial cells Yes Cronauer, M.V. et al. (1999) Am. J. Pathol. 155, 1977–84. PubMed ID #10595927.
Human cerebrovascular tissue Yes Cronauer, M.V. et al. (1999) Am. J. Pathol. 155, 1977–84. PubMed ID #10595927.
Human adenoma tissue Yes Jaquet, P. et al. (2000) J. Clin. Endocrinol. Metab. 85, 781–92. PubMed ID #10690891.
Human hair follicle Yes Use fresh samples and at least 10 or more hair roots. Align the hairs, and cut the root end from the hair and quickly process using the SV RNA Isolation System. The amount of RNA is very small and can generally only be detected by RT-PCR. Contact Promega Technical Services for more information.
Ferret aorta Yes Gangopadhyay, S.S. et al. (2002) Biochem. J. 372, 347–57. PubMed ID #12603201.
Feline oral mucosa Yes Harley, R. et al. (1999) Clin. Diag. Lab. Immunol. 6, 471–8. PubMed ID #10391845.
Sample Type Promega External Comments
Plant tissues A general protocol for total RNA isolation from plant tissue is provided in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources are listed below.
Tomato leaf Yes Follow the standard protocol for plant tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Shenoi, H. and Kephart, D. Titration of tomato leaf material used as starting material for SV total RNA isolation. eNotes Kobs, G. (1998) Promega Notes 68, 28–31.
Arabidopsis thaliana leaf Yes Yes Follow the standard protocol for plant tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Technically Speaking: SV Total RNA Isolation System. Promega Notes 68, 16–9.Caldelari, D. et al. (2001) Plant Physiol. 126, 1416–29.PubMed ID #11500541.
Arabidopsis thaliana flower, stem and root Yes Caldelari, D. et al. (2001) Plant Physiol. 126, 1416–29. PubMed ID #11500541.
Tobacco leaf Yes Follow the standard protocol for plant tissue in the SV Total RNA Isolation System Technical Manual #TM048.
Sugar beet 14-day seedling, cotyledon, hypocotyl, 46-day plant and 90-day plant) Yes Fowler, M.R. et al. (2000) J. Exp. Bot. 353, 2125–6. PubMed ID #11141186.
Chicory taproot, etiolated heads, leaf, and green and etiolated seedlings Yes Fowler, M.R. et al. (2000) J. Exp. Bot. 353, 2125–6. PubMed ID #11141186.
Three-day-old mung bean hypocotyls Yes Leiter, H. et al. (1999) J. Biol. Chem. 274, 21830–9. PubMed ID #10419500.
Pea seedling leaf Yes Skaf, J.S. et al. (2000) J. Gen. Virol. 81, 1103–9. PubMed ID #10725438.
Plant tissues A general protocol for total RNA isolation from plant tissue is provided in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources are listed below.
Tomato leaf Yes Follow the standard protocol for plant tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Shenoi, H. and Kephart, D. Titration of tomato leaf material used as starting material for SV total RNA isolation. eNotes Kobs, G. (1998) Promega Notes 68, 28–31.
Arabidopsis thaliana leaf Yes Yes Follow the standard protocol for plant tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Technically Speaking: SV Total RNA Isolation System. Promega Notes 68, 16–9.Caldelari, D. et al. (2001) Plant Physiol. 126, 1416–29.PubMed ID #11500541.
Arabidopsis thaliana flower, stem and root Yes Caldelari, D. et al. (2001) Plant Physiol. 126, 1416–29. PubMed ID #11500541.
Tobacco leaf Yes Follow the standard protocol for plant tissue in the SV Total RNA Isolation System Technical Manual #TM048.
Sugar beet 14-day seedling, cotyledon, hypocotyl, 46-day plant and 90-day plant) Yes Fowler, M.R. et al. (2000) J. Exp. Bot. 353, 2125–6. PubMed ID #11141186.
Chicory taproot, etiolated heads, leaf, and green and etiolated seedlings Yes Fowler, M.R. et al. (2000) J. Exp. Bot. 353, 2125–6. PubMed ID #11141186.
Three-day-old mung bean hypocotyls Yes Leiter, H. et al. (1999) J. Biol. Chem. 274, 21830–9. PubMed ID #10419500.
Pea seedling leaf Yes Skaf, J.S. et al. (2000) J. Gen. Virol. 81, 1103–9. PubMed ID #10725438.
Plant tissues A general protocol for total RNA isolation from plant tissue is provided in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources are listed below.
Tomato leaf Yes Follow the standard protocol for plant tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Shenoi, H. and Kephart, D. Titration of tomato leaf material used as starting material for SV total RNA isolation. eNotes Kobs, G. (1998) Promega Notes 68, 28–31.
Arabidopsis thaliana leaf Yes Yes Follow the standard protocol for plant tissue in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Technically Speaking: SV Total RNA Isolation System. Promega Notes 68, 16–9.Caldelari, D. et al. (2001) Plant Physiol. 126, 1416–29.PubMed ID #11500541.
Arabidopsis thaliana flower, stem and root Yes Caldelari, D. et al. (2001) Plant Physiol. 126, 1416–29. PubMed ID #11500541.
Tobacco leaf Yes Follow the standard protocol for plant tissue in the SV Total RNA Isolation System Technical Manual #TM048.
Sugar beet 14-day seedling, cotyledon, hypocotyl, 46-day plant and 90-day plant) Yes Fowler, M.R. et al. (2000) J. Exp. Bot. 353, 2125–6. PubMed ID #11141186.
Chicory taproot, etiolated heads, leaf, and green and etiolated seedlings Yes Fowler, M.R. et al. (2000) J. Exp. Bot. 353, 2125–6. PubMed ID #11141186.
Three-day-old mung bean hypocotyls Yes Leiter, H. et al. (1999) J. Biol. Chem. 274, 21830–9. PubMed ID #10419500.
Pea seedling leaf Yes Skaf, J.S. et al. (2000) J. Gen. Virol. 81, 1103–9. PubMed ID #10725438.
Sample Type Promega External Comments
Cultured cells (Established cell lines are not listed.) Yes A general protocol for total RNA isolation from mammalian cultured cells is provided in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources are listed below.
Primary rat endothelial cells Yes Thum, T., Haverich, A. and Borlak, J. (2000) FASEB J. 14, 740–51. PubMed ID #10744630.
Fibroblasts derived from mouse embryonic stem cells Yes Zhang, S., Day, I.N.M. and Ye, S. (2001) Physiol. Genomics 5, 187–92. PubMed ID #11328964.
Primary human coronary artery endothelial cells Yes Zhang, S., Day, I.N.M. and Ye, S. (2001) Physiol. Genomics 5, 187–92. PubMed ID #11328964.
Sample Type Promega External Comments
Bacteria A general protocol for total RNA isolation from Gram-positive and Gram-negative bacteria is provided in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources are listed below.
Escherichia coli Yes Yes Follow the standard protocol for bacteria in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Kobs, G. (1998) Promega Notes 68, 28–31. Ohmichi, T., Maki, A. and Kool, E.T. (2002) Proc. Natl. Acad. Sci. USA 99, 54–9. PubMed ID #11752404.
Bacillus subtilis Yes Yes Follow the standard protocol for bacteria in the SV Total RNA Isolation System Technical Manual #TM048. Additional resources: Kobs, G. (1998) Promega Notes 68, 28–31. Colombo, M.L. et al. (2004) Antimicrob. Agents Chemother. 48, 484–90. PubMed ID #14742199.
Vibrio vulnificus Yes Chen, Y.C. et al. (2004) Infect. Immun. 72, 629–36. PubMed ID #14742502.
Salmonella enterica Yes Kelly, A. et al. (2004) Microbiology 150, 2037–53. PubMed ID #15256548. Klein, J.R., Fahlen, T.F. and Jones, B.D. (2000) Infect. Immun. 68, 3368–76. PubMed ID #10816487.
Streptomyces coelicolor Yes The biomass from pregerminated Streptomycetes coelicolor spores was collected, frozen in liquid nitrogen and crushed by mortar and pestle. The resulting powder was processed to extract RNA using the TRIzol® reagent, and the aqueous phase was treated with DNase I. SV RNA Dilution Buffer (375µl) was added to the nucleic acid solution, and samples were mixed and centrifuged. The resulting supernatant was mixed with 250µl of cold 96% ethanol, loaded on an SV spin column and centrifuged for 1 minute. The column was then processed as described in the SV Total RNA Isolation System Technical Manual #TM048. Van Keulen G. et al. (2004) J. Microbiol. Methods 58, 139–42. PubMed ID #15177913.
Bacillus thuringiensis Yes Lee, M.K. et al. (2003) Appl. Environ. Microbiol. 69, 4648–57. PubMed ID #12902253.
Alteromonas Yes Orikoshi, H. et al. (2003) J. Bacteriol. 185, 1153–60. PubMed ID #12562783. Tsujibo, H. et al. (2000) Appl. Environ. Microbiol. 66, 3778–83. PubMed ID #10966390.
Pseudomonas aeruginosa Yes After resuspension in a lysis buffer (50mM Tris-HCl [pH 7.5], 150mM NaCl), the cell pellet was subjected to French press, and 100μl aliquots were processed with the SV Total RNA Isolation System as directed by the manufacturer. Cambronne, E.D. and Schneewind, O. (2002) J. Bacteriol. 184, 5880–93. PubMed ID #12374821.
Streptococcus pneumoniae Yes Brown, J.S. et al. (2002) Infect. Immun. 70, 4389–98. PubMed ID #12117949.
Mycobacterium smegmatis Yes Milano, A. et al. (2001) J. Bacteriol. 183, 6801–6. PubMed ID #11698368.
Klebsiella pneumoniae Yes Yigit, H. et al. (2001) Antimicrob. Agents Chemother. 45, 1151–61. PubMed ID #11257029.
Rhizobium leguminosarum Yes Lee, H.Y., An, J.H. and Kim, Y.S. (2000) Eur. J. Biochem. 267, 7224–9. PubMed ID #111064359.
Pseudomonas putida Yes Cowles, C.E., Nichols, N.N. and Harwood, C.S. (2000) J. Bacteriol. 182, 6339–46. PubMed ID #11053377. Grimm, A.C. and Harwood, C.S. (1999) J. Bacteriol. 181, 3310–6. PubMed ID #10322041.
Rhodopseudomonas palustris Yes Pelletier, D.A. and Harwood, C.S. (2000) J. Bacteriol. 182, 2753–60. PubMed ID #107815431.
Neiserria meningitidis Yes Deghmane, A.E. et al. (2000) EMBO J. 19, 1068–78. PubMed ID #106989471.
Lactobacillus helveticus Yes Pederson, J.A. et al. (1999) J. Bacteriol. 181, 4592–7. PubMed ID #10419958.
Simkania negevensis ZT Yes Everett, K.D. et al. (1999) J. Bacteriol. 181, 4734–40. PubMed ID #10438738.
Chlamydia trachomatis Yes Everett, K.D. et al. (1999) J. Bacteriol. 181, 4734–40. PubMed ID #10438738.
Sample Type Promega External Comments
Saccharomyces cerevisiae Yes A protocol is provided in the SV Total RNA Isolation System Technical Manual #TM048. Additional resource: Kobs, G. (1998) Promega Notes 68, 28–31.
Gigaspora margarita Yes Lanfranco, L. et al. (2002) Plant Physiol. 130, 58–67. PubMed ID #12226486.
Trichomonas vaginalis Yes Coombs, G.H. et al. (2004) J. Biol. Chem. 279, 5249–56. PubMed ID #14630923.
Pacific oyster larvae Yes Steele, S. and Thorndyke, M. (1999) Promega Notes 71, 2–4.
Cartilaginous fish brain Yes Pederson, J.A. et al. (1999) J. Bacteriol. 181, 4592–7. PubMed ID #10419958.
Zebrafish gut Yes Crosnier, C. et al. (2005) Development 132, 1093–104. PubMed ID #15689380.
Drosophila larva tissue Yes Petryk, A. et al. (2003) Proc. Natl. Acad. Sci. USA 100, 13773–8. PubMed ID #14610274.
Frog liver Yes Saeed-Kothe, A. and Powers-Lee, S.G. (2003) J. Biol. Chem. 278, 26722–6. PubMed ID #12738780.
Frog gonads and mesonephroi Yes Ogata, M. et al. (2003) Genetics, 164, 613–20. PubMed ID #12807781.
Chicken embryonic heart, brain and skeletal muscle Yes Agarkova, I. et al. (2000) J. Biol. Chem. 275, 10256–64. PubMed ID #10744711.
Venom sacs and glands from Naja Atra (Asian cobra) Yes Lou, X. et al. (2004) J. Biol. Chem. 279, 39094–104. PubMed ID #15252034.
Microcystis aeruginosa Yes Kaebernick, M. et al. (2002) Appl. Environ. Microbiol. 68, 449–55. PubMed ID #11823177.
Sample Type Promega External Comments
In vitro transcribed RNA Yes M. Raynor, J.O. and Butler, K.D. (2004) Promega Notes 86, 15–7.
Human immunodeficiency virus Yes Personal communication. Contact Promega Technical Services for more information.
RNA from soil Yes Personal communication. Contact Promega Technical Services for more information.

Protocols

Complete Protocol

SV Total RNA Isolation System Protocol


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Quick Protocol

SV Total RNA Isolation System Quick Protocol FB025

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Specifications

What’s in the box?

Item - Catalog number: Z3101 Part # Size Available Separately
Collection Tubes A130E 2 × 5/pk
Nuclease-Free Water P119A 1 × 1,250μl View Product
Elution Tubes Z200A 2 × 5/pk
RNA Lysis Buffer (RLA) Z305B 1 × 10ml View Product
RNA Dilution Buffer (RDA) Z306B 1 × 4ml
RNA Wash Solution (RWA) Z309B 1 × 11.8ml View Product
DNase Stop Solution (DSA) Z312A 1 × 5.3ml
Yellow Core Buffer Z317A 1 × 2.5ml
MnCl2, 0.09M Z318A 1 × 250μl
DNase I (lyophilized) Z358B 1 × 1 vial
Spin Columns Z503B 2 × 5/pk
Beta-Mercaptoethanol Z559A 1 × 900μl

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

What’s in the box?

Item - Catalog number: Z3100 Part # Size Available Separately
Collection Tubes A130D 2 × 25/pk
Nuclease-Free Water P119E 1 × 13ml View Product
Elution Tubes Z200B 2 × 25/pk
RNA Lysis Buffer (RLA) Z3051 1 × 50ml View Product
RNA Dilution Buffer (RDA) Z306A 1 × 20ml
RNA Wash Solution (RWA) Z3091 1 × 58.8ml
DNase Stop Solution (DSA) Z312A 1 × 5.3ml
Yellow Core Buffer Z317A 1 × 2.5ml
MnCl2, 0.09M Z318A 1 × 250μl
DNase I (lyophilized) Z358A 1 × 1 vial
Spin Columns Z503C 2 × 25/pk
Beta-Mercaptoethanol Z559A 1 × 900μl

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

What’s in the box?

Item - Catalog number: Z3105 Part # Size Available Separately
Collection Tubes A130A 5 × 50/pk
Nuclease-Free Water P119C 2 × 25ml View Product
Elution Tubes Z200C 5 × 50/pk
RNA Lysis Buffer (RLA) Z305E 1 × 250ml View Product
RNA Dilution Buffer (RDA) Z306C 1 × 96ml
RNA Wash Solution (RWA) Z309D 1 × 206ml View Product
DNase Stop Solution Z312D 1 × 26.5ml
Yellow Core Buffer Z317C 1 × 11ml
MnCl2, 0.09M Z318B 2 × 750μl
Spin Columns Z503D 5 × 50/pk
Beta-Mercaptoethanol Z559A 1 × 900μl

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions