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Old 04-22-2015, 12:02 PM   #4
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Location: New England

Join Date: Jun 2012
Posts: 200

Hopefully, the 1st step PCR will work perfectly the same with tailed primers! In reality, this is probably not the case, but if you can't get the PCR to work properly with just the locus primers, it's not going to work with the tailed primers. So we recommend testing with the cheaper locus primers to make sure it works - in our experience, once users get the locus primers to work, the tailed primers also work. I also discourage using gel extraction as a way to get around multiple bands from the first PCR - the second PCR is pretty robust, but I haven't had success using it on gel-extracted samples.

For the second round of PCR, you should use the PCR conditions outlined by Illumina. We actually use different conditions, since we "developed" the method based on a slide show from Illumina prior to them issuing the official protocol.

I've had a look online for publications from our investigators but there don't seem to be any yet - most of the runs were within the last year so it may be a bit early for papers.
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