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sequencing the pcr product


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15 replies to this topic

#16 Trof

Trof

    Brain on a stick

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Excellent

Posted 23 February 2009 - 03:22 AM

Can we get clean sequencing results without purifying it? I thought the excess primers in the PCR products would also "participate" in the sequencing reaction and synthesizing sequence, for example, from the other end, and thus causing the background noise...

Many things can cause background noise, not HPLC-cleaned primers, unpurified cycle sequencing,...sometimes you can't even figure why, so I wouldn't try to put unpurified product to the reaction and expect no background. It's too expensive to let it fail on that.

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