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Old 06-15-2009, 02:18 AM   #2
james hadfield
Cambridge, UK
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Location: Cambridge, UK

Join Date: Feb 2008
Posts: 221

I could not see your initial library on the gel, unlikely I know but running it alongside as a positive control might be helpful. Were all the other libraries the same 350bp size and did you run the PhiX as another control?

It looks like you are using a SYBR based approach which will be confounded by the average insert size when trying to determine the number of molecules present as opposed to the amount of DNA. A TaqMan assay would be more reliable and I think this was proposed in the Quail et al publication.

We are almost there on getting TaqMan QC ito our protocol, but it has been challenging to make it as robust as possible.
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