RNA contamination PCR - RNA contamination PCR (Apr/02/2003 )
i ve some RNA contamination in PCR ( to amplify a gene)
expected product is amplified to a very less extent even after 35 cycles
can U help me?
first of all u can add RNAse A 10mg/ml (3 ul for 100 ul solution). second RNA contamination do not disturbe the PCR reaction at all. u need dsDNA to amplified the gene u want. if u can see the desired band u can use gel purification kit to isolate the band and then do PCR while the purified band will be now the template.
p.s. be sure that what u see is RNA and not nonspesific bands (look like a smir) as a result of low TM condition.