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What’s wrong with my NTC - (Sep/09/2010 )

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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-patchbear-

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

-ivanbio-

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 ?

-patchbear-