Some problems in RT-PCR - Biology Forum

last updated by

tranjo 18 years, 5 months ago

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- May 30, 2006 at 11:57 pm #4917
  schmyly
  Participant
  I’m carrying on RT-PCR and met some problems. Could anyone help me?
  1. What problem will be caused by DNA contamination of RNA in RT-PCR?
  2. What’s the optimal Mg2+ concentration in the PCR system using pfu
  polymerase?
- June 3, 2006 at 4:01 pm #49427
  tranjo
  Participant
  The problem of DNA contamination depends on your purpose. Quantitative measures like real-time PCR can’t be done with DNA contamination when you do RT – it skews the results. If you want to amplify some known coding sequences, it shouldn’t be a big deal. You can order most full-length clones, however.
  There is no real optimal Mg2+ for PCR. Pfu will work over a wide-range of Mg2+ concentrations. It will not work with excessive amounts of Mg2+ or anything else crazy.
  Best, J
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