Long distance RT-PCR

Technique tips for long-distance RT-PCR:

1. Extract poly-A mRNA using oligo-(dT)25 dynabeads. Analyze mRNA quality before RT reaction (Northern blot).

2. Perform RT reaction using SuperScriptII or SuperScriptIII reverse transcriptase per manufacturer's instruction.

3. Follow long PCR technology guideline for amplification of extra long DNA template. (commercial kits available from Roche and Life technologies).

Related papers:

J Virol Methods. 2006 Jun;134(1-2):197-204. A long distance RT-PCR able to amplify the Pestivirus genome.

Last update 01-May-2007, Rating n/a of 0 votes.