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RT-PCR band from divergent genes troubleshooting

RT-PCR troubleshooting

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#1 cpshutterbug



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Posted 01 March 2012 - 09:25 AM

Hi all,

I have a question related to problems with an RT-PCR. Basically, I have three controls. One is the no RT control, and the others are the positive gDNA control, and a negative control involving primers designed between two divergently transcribed genes. When I run the RT-PCR, what I see is no band for the NRT control, yet there is a faint band in the RT reaction for the divergently transcribed genes. I tried analyzing a different set of divergently transcribed genes with the same faint band as a result. Also, I left spaces between wells on the agarose gel to see if there was any carryover from another well. No changes there. Has anyone seen this before? Any suggestions on how to solve this problem or as to what could be causing it?

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