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RNA contamination PCR


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#1 visu_bio

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Posted 02 April 2003 - 10:14 AM

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?
visu

#2 yossi.b

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Posted 04 April 2003 - 12:01 AM

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.
good luck

yossi




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