I have extracted RNA from tumor tissue sample and wondering whether the basal level of p53 expression enough to do RT PCR amplification using p53 transcript specific primers?
Thank you
Dear Vahinpiriya,
May I ask you why you want to do RT by specific primers?
I think it is better to use Random hexamer and Oldio dT for cDNA synthesis then measure the expression level of your gene of interest.
Normally p53 expression is low in tumor samples but not that much that you would not detect it.
Hi Elyas,
I actually wanted to do cDNA sequencing. So after cDNA synthesis, I wanted to amplify cDNA of p53 using specific primers
There are enough transcripts for that purpose.
You can perform your cDNA synthesis by general primers (Random hexamer and oligo dT) and then amplificate p53 by specific primers and PCR.
Run your PCR product on the agarose gel. If you got just one sharp band, without smear and primer dimer, it is suitable for sequencing.
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