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t4taylor's Content

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#120325 Direct PCR for Plants (Arabidopsis)

Posted by t4taylor on 25 September 2011 - 09:56 AM in PCR, RT-PCR and Real-Time PCR

Hi everyone,

Our lab used to use Sigma’s Extract-N-Amp™ Plant direct PCR kit for quickly genotyping large numbers of plants. It always worked amazingly well – unfortunately, the buffers for this kit have run out and are now too expensive to replace (£1,410!).

I’ve been searching for an alternative direct PCR plant protocol for a while now, and sadly, none of the protocols I’ve found work reliably enough to use on a regular basis.

I was wondering if anyone had any direct PCR plant protocols that they use and are happy with?

Common themes for the protocols seem to involve a sodium hydroxide extraction, heating the sample to around 95°C and then a neutralisation or dilution step.

Any advice anyone?

Many thanks,

Ben




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