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pcr trouble!!! heeeeeellllpppppp!!

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#1 MaggieRoara



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Posted 14 October 2009 - 11:14 PM

I am having a really difficult time with troubleshooting a PCR reaction thats not working. I am not seeing any PCR bands. I have narrowed it down to the problem either being because of the primers or the plasmid template. Is it possible for thereto be something wrong with my template. I checked it on a gel, there's a band at the correct place i.e. the miniprep to get the template was successful.

#2 phage434



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Posted 15 October 2009 - 03:45 AM

Step 1: retry the PCR using 10x and 100x diluted template plasmid
Step 2: redesign your primers, perhaps with some help. Primer3 is a good program, and the IDT primer analyzer is good (idtdna.com)

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