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False positive - help


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3 replies to this topic

#1 krazyhorz

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Posted 13 June 2014 - 01:30 PM

Hi.
As a newcomer to the forum i would greatly appreciate any help anyone could offer. I am running PCR to amplify approx 150bp sequence. I am getting a strong band on gel for the pos control isolate but other isolates, which I know should be not be positive, are giving fainter but definite bands at same 150bp position as pos control. However, my negative control is negative. Can anyone shed any light on what could be going on? Thanks

#2 merlav

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Posted 16 June 2014 - 04:58 AM

Or those samples aren't negative as you thought or the amplification band that you are seen isn't the target and are unspecify amplification


Science without religion is lame, religion without science is blind.
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I am among those who think that science has great beauty. A scientist in his laboratory is not only a technician: he is also a child placed before natural phenomena which impress him like a fairy tale.
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#3 superice

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Posted 25 June 2014 - 07:30 AM

change the primer you used, it not suitable for your experiment. you could get the primer sequence from   http://pga.mgh.harvard.edu/primerbank/   or re-design them



#4 superice

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Posted 25 June 2014 - 07:30 AM

change the primer you used, it not suitable for your experiment. you could get the primer sequence from   http://pga.mgh.harvard.edu/primerbank/   or re-design them






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