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Old 03-31-2011, 08:42 AM   #9
pmiguel
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Location: Purdue University, West Lafayette, Indiana

Join Date: Aug 2008
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Yes you are talking about a "bottomed out" library. You start with a certain amount of DNA. For SureSelect, I don't know how much. But rule of thumb: 1 ug of DNA fragmented to average 1 kb size comprises a trillion (10^12) molecules. Probably you are closer to 10 trillion initially because fragmentation would be down to closer to 100 bp?

Then you have a series of steps that winnows the total number of real fragments down. Each is a bottle-neck. How narrow is the bottle-neck? Often this is hard to assess--especially when PCR is being deployed. 10 cycles of PCR potentially can amplify the amount of DNA you have 1000-fold. That is, picograms become nanograms. Or, one million molecules becomes one billion. But, obviously the billion will all be copies of the original million. Yes, something to be concerned about.

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Phillip
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