Home | Molecular Biology | Electrophoresis & Nucleic Acid Purification | Nucleic Acid Purification | Spin column-based isolation

GeneJET RNA Purification Kit


Silica membrane-based total RNA purification from mammalian blood, cell cultures, and tissues as well as insect, yeast, and bacteria. Provided in a convenient spin-column format.
  
Loading...

The Thermo Scientific 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.

Highlights

  • 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, and 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
  • 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
Table 1. | Typical RNA yields from various sources.
Source Quantity Yield, µg
Mouse heart 20 mg 10 to 15
Mouse muscle 30 mg 8 to 10
Mouse lung 30 mg 25 to 30
Mouse kidney 30 mg 25 to 30
Mouse liver 30 mg 60 to 65
Mouse spleen 5 mg 10 to 15
Bacillus pumilis cells 1 x 109 cells 15 to 20
Escherichia coli cells 1 x 109 cells 25 to 30
HeLa cells 5 x 106 cells 35 to 40
Jurkat cells 5 x 106 cells 40 to 50
Cos-7 cells 1 x 106 cells 20 to 25
Saccharomyces cerevisiae cells 4 x 108 cells 150 to 160
  
Storage ConditionAmbient
HazardousNo
Quality Control
  • The 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.
Overview of purification procedure

Overview of purification procedure

Overview of purification procedure

Figure 1 | Overview of purification procedure.


Total RNA from various sources purified using the GeneJET RNA Purification Kit

Total RNA from various sources purified using the GeneJET RNA Purification Kit

Total RNA from various sources purified using the GeneJET RNA Purification Kit

Figure 2 | 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


Yields of total RNA purified using the GeneJET RNA Purification Kit and kits of competitors

Yields of total RNA purified using the GeneJET RNA Purification Kit and kits of competitors

Yields of total RNA purified using the GeneJET RNA Purification Kit and kits of competitors

Figure 3 | Yields of total RNA purified using the GeneJET RNA Purification Kit and kits of competitors.


References

  1. P. Chomczynski, N. Sacchi, Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal. Biochem. 162(1), 156-159 (April 1987).

  2. R. Boom, C. J. A. Sol, M. M. M. Salimans, C. L. Jansen, P. M. E. W. Dillen, J. van der Noordaa, Rapid and simple method for purification of nucleic acids. J. Clin. Microbiol. 28(3), 495–503 (March 1990).

Protocols and Product Manuals

pdf   GeneJET RNA Purification Kit - Product Information