RT-PCR without RNA isolation.

@article{Klebe1996RTPCRWR,
  title={RT-PCR without RNA isolation.},
  author={Robert J. Klebe and George M Grant and Althea M. Grant and Miguel Angel Rodriguez Garcia and Troy A. Giambernardi and Graham P Taylor},
  journal={BioTechniques},
  year={1996},
  volume={21 6},
  pages={
          1094-100
        }
}
Reverse transcription-PCR (RT-PCR) has traditionally required time-consuming RNA extraction and purification. This report demonstrates that one can completely avoid the RNA extraction step in RT-PCR by basing the comparison of samples on cell number rather than micrograms of total RNA. A new method for lysing cells while preserving RNA is described. RT-PCR is carried out (i) by rapidly freezing cells in the presence of ribonuclease inhibitor (RNase inhibitor) plus dithiothreitol and (ii) by… CONTINUE READING

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