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GeneJET™ RNA Purification Kit
- Product information
-
Overview of purification procedure.
- Features
- Universal – can be used for both cell and tissue samples from a wide range of sources: mammalian blood, mammalian cell cultures, mammalian tissues, insect, yeast, bacteria.
- Efficient – high yields of total RNA.
- Fast – 15 minutes after lysis step.
- Pure – an A260/280 ratio > 1.9.
- Convenient – spin columns are capped and assembled with collection tubes.
- Applications
- Fast extraction of high purity RNA suitable for all conventional molecular biology procedures, including:
– RT-PCR,
– RT-qPCR,
– Northern blotting,
– nuclease protection assay.
- Fast extraction of high purity RNA suitable for all conventional molecular biology procedures, including:
DescriptionThe GeneJET™ RNA Purification Kit is a simple and efficient system for purification of total RNA from mammalian cultured cells, tissue, human blood cells, bacteria and yeast.The kit utilizes a silica-based membrane technology in the form of a convenient spin column, eliminating the need for tedious cesium chloride gradients, alcohol precipitation or toxic phenol-chloroform extractions.
RNA molecules longer than 200 nucleotides can be isolated with the GeneJET™ RNA Purification Kit in 15 minutes after the lysis step. The high-quality purified RNA can be used in a wide range of downstream applications including RT-PCR, RT-qPCR, Northern blotting and other RNA-based analyses.
Quality ControlThe GeneJET™ RNA Purification Kit is qualified by isolating total RNA from 20 mg of mouse muscle or heart following the described protocol. The purified RNA has an A260/280 ratio of ≥1.9. The RNA integrity numbers (RIN) of RNA purified from mouse muscle and heart are >7 and >8, respectively. Functional quality of the purified RNA is evaluated by RT-PCR of mouse dystrophin RNA resulting in a 13.3 kb PCR product.
Components- Proteinase K
- Lysis Buffer
- Wash Buffer 1 (concentrated)
- Wash Buffer 2 (concentrated)
- Water, nuclease-free
- GeneJET™ RNA Purification Columns (pre-assembled with Collection Tubes)
- Collection Tubes (2 ml)
- Collection Tubes (1.5 ml)
- Detailed Protocol
Figure 1. Total RNA from various sources purified using the GeneJET™ RNA Purification Kit.
1 µg of isolated total RNA was analyzed by agarose gel (1%) electrophoresis.
M – RiboRuler™ High Range RNA Ladder (#SM1821)
1 – mouse liver
2 – mouse spleen
3 – HeLa cells
4 – Cos-7 cells
5 – Saccharomyces cerevisiae
6 – Escherichia coli DH5α
Figure 2. Yields of total RNA purified using the GeneJET™ RNA Purification Kit and kits of competitors.
Table 1. Typical RNA yields from various sources.Source Quantity Yield, µg Mouse heart 20 mg 10-15 Mouse muscle 30 mg 8-10 Mouse lung 30 mg 25-30 Mouse kidney 30 mg 25-30 Mouse liver 30 mg 60-65 Mouse spleen 5 mg 10-15 Bacillus pumilis cells 1x109 cells 15-20 Escherichia coli cells 1x109 cells 25-30 HeLa cells 5x106 cells 35-40 Jurkat cells 5x106 cells 40-50 Cos-7 cells 1x106 cells 20-25 Saccharomyces cerevisiae cells 4x108cells 150-160
Patents, Licenses, Trademarks - Protocols & recommendations
- References
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- Chomczynski, P. and Sacchi. N., Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction, Anal. Biochem., 162, 156-159, 1987.
- Boom, R., Sol C.J.A., Salimans, M.M.M. Jansen, C.L. Dillen, P.M.E.W. and van der Noordaa J., Rapid and simple method for purification of nucleic acids, J. Clin. Microbiol., 28, 495–503, 1990.
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- RevertAid™ Premium Reverse Transcriptase
- Maxima® Reverse Transcriptase
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- Maxima® First Strand cDNA Synthesis Kit for RT-qPCR
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