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Old 06-04-2014, 08:01 AM   #14
pmiguel
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Location: Purdue University, West Lafayette, Indiana

Join Date: Aug 2008
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Quote:
Originally Posted by nucacidhunter View Post
The result will be underestimation of library concentration because only a portion of it is quantified. Other issue would be differences in amplification efficiency of standards (~400 bp) with library. I have verified this by running QPCR product on DNA Chip and comparing its size to input amplicon size in large insert libraries. I have found that the amplicon peak of QPCR is substantially lower than actual input DNA.
Okay, let's see the before and after (q)PCR DNA Chips.

Also, wasn't it necessary to remove the SYBR green, etc. from the qPCR reaction prior to running the chip? What method did you use?

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Phillip
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