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Old 08-18-2015, 11:10 PM   #1
Shiwali Goyal
Junior Member
Location: India

Join Date: Aug 2015
Posts: 9
Default Exome sequencing troubleshootings

Im doing whole exome sequencing on Ion torrent platform. I started with 1.4microgram/microliter of stock DNA. The fragmentation of the DNA was done correctly as I can see the smear on E-gel near 130-150basepair. Next I proceed with the adapter ligation of the library and then to the size selection step of the library preparation. On library preparation step I picked up the library of 210 basepair size. Then I proceed with the size selected library amplification. When I checked the Qubit concentration of my amplified library, it was below 4 ng/microliter (required 500ng). This concentration must be near 10 or 11 ng/microlitre. Can anyone tell me where the flaw is in this whole experiment?
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