PCR not consistent

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elynn
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PCR not consistent

Post by elynn » Wed Dec 05, 2012 9:12 am

Dear everyone,

I have a problem with my PCR. Its not consistent. Sometimes i got bands on my gel and sometimes i don't get bands. however the mixture and pcr protocol is the same. I hv check my DNA and primers and they're in good conditions. Is it normal to have this kind of problem? Can anyone suggest some solutions?

Thanks

emadsadeghian
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Post by emadsadeghian » Fri Jan 04, 2013 12:30 pm

have u tried adding dmso?

gislab03
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Post by gislab03 » Sat May 04, 2013 2:16 pm

Do you include positive control for every run and does the positive control give appropriate band? And agarose gel is also important in visualising the results. You should use freshly prepared agarose and ethidium bromide solution.

cathbr
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Post by cathbr » Tue Jun 18, 2013 4:59 pm

What about the template that you are using? what is it? genomic DNA, plasmid,...? Are you sure that you are always using the same concentration and that the template quality is good and not degraded? and what kind of enzyme do you use?

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