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What’s wrong with my NTC


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#1 patchbear

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Posted 09 September 2010 - 06:36 AM

My NTC always has a band which is the same size as my product(about 150bp). Maybe contamination……but I changed everything for PCR, including master mix , water and primer……I can still see the band ,it has been for several weeks ,my god ,help……

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Edited by patchbear, 09 September 2010 - 06:37 AM.


#2 ivanbio

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Posted 09 September 2010 - 07:18 AM

If you've already done everything under the sun to clean up your reagents and still get this band in your NTC, then there are two possible things you can do:

1. Buy new primers. This background band could well be your primers coming together in some weird structure that happens to run at this size
2. Design a new assay. Having a band at 150 bp is very close to where all the issue arise. Design an assay that gives you a larger band, at least 300 bp.

My two cents

Ivan
Carlsbad, CA

#3 patchbear

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Posted 09 September 2010 - 04:19 PM

thank you ivanbio
but what's the size of the pds?there are two bands in my NTC,one is primer dimer, the other is weird structure ?




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