RT-PCR

Erwin Pauws & Carrie Ris ris at eland.amc.uva.nl
Mon Feb 26 15:09:52 EST 1996


Excess amount of cDNA in your PCR might inhibit the amplification due to 
contamination, that's why lowering the amount can better your PCR-results.
Use as little cDNA as possible, you can check this by amplifying some 
house-keeping gene with a titration of cDNA, always next to an -RT (cDNA
synthesis negative control without any RT-enzym; this to check for DNA
contamination in your RNA.
Also, first-strand synthesis using RANDOM HEXAMERS instead of OLIGO-dT gives
better results (more cDNA copies).




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