RT-PCR suggestion??

S M Shamsul Islam @S-M-Shamsul-Islam

15 August 2016 1 3K Report

I am doing RT-PCR from human blood sample with the hBeta actin.

But the bands (in different groups) showing different expression.

some are high some r low. In case of beta actin it should show, same expression of all bands right???

what should be the trouble shooting

Srinivas H Gowda

different loading volume might give this pattern. rept once even then it comes the same then it could be that the RT is not optimal in disease 1 sample or the RNA quality is low. if you have sufficient RNA can do RNA gel electrophrosis to know the quality of RNA. It is rarely that actin housekeeping gene changes that significantly, if that is the case you need to change the housekeeping gene to some other one.

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