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Simple and Rapid Isolation of Plant RNA,,, Using the Eppendorf Perfect RNA, Eukaryotic, Maxi Kit

ontrol without RNA. Discussion

The Eppendorf Perfect RNA Kit for eukaryotic cells is well suited for the isolation of intact total RNA from plants. Within 2.5 hours, highly purified, high molecular weight total RNA can be obtained from various source materials such as leaves, flowers, pods, and plant cell cultures. The method is efficient and eliminates exposure to hazardous organics. Conventional phenol/chloroform extraction with subsequent ethanol precipitation requires almost twice the time. Also, the buffer supplied with the kit significantly reduces the risk of RNase contamination during isolation. 180-800 g RNA can be isolated from 1-5 g plant material. The use of more than 5 g source material does not result in a pronounced increase in yield. When plant cell cultures were used as the starting material, the RNA yield was almost doubled compared to the plant material. The measurement of the A260/280 ratios confirmed the reproducible extraction of highly pure, intact, total RNA. RNA isolated with the Perfect RNA Kit can be directly used in demanding subsequent reactions like RT-PCR.

References

  • Chomczynski P, Sacchi N., Anal Biochem 1987 Apr;162(1):156-9, Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.
  • Chirgwin, J.M., et al. (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochem. 18(24): 5294-5299.
  • Maniatis, T., Fritsch, E.F., and Sambrook, J. (1982) Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY. pp.188-
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