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Old 04-22-2017, 06:24 PM   #2
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,230
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Only P2 adapter in ddRAD has short non-complementary overhang so its effect on anomalous migration of ligated fragments in microfluidics separation devices such as Bioanalyser is less pronounced in comparison with standard forked adapters. This effect also is minor on agarose gels and causes a small shift in size-selection on Pippin devices. You need to specify size selection range considering added adapter length. If your library needs precise size range you may need to do a trial to adjust for the size shift.


Advantages of size-selection in ddRAD before PCR:
1- Low mass of DNA overcoming using multiple lanes for size-selection
2- Less bias in PCR amplification and therefore more uniform tag coverage because of uniform fragment sizes
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