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PCR help

PCR help - PCR - Polymerase Chain Reaction Forum

PCR help - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 11-25-2011, 09:53 AM
Pipette Filler
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Default PCR help



Hi,

So i have designed primers using primer premier 6.0 and i have tried to amplify a cDNA fragment of my gene so that i can clone it into a pGMET vector and synthesis riboprobes to analyze the expression patterning using whole-mount.

However, after sequencing and with a blast search, i realized the cDNA fragment i amplified was not the one i was interested in.

I was wondering what the cause of this non-specific amplification could be. I would be grateful for your suggestions.
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