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Old 11-29-2012, 05:29 AM   #1
miraiama
Junior Member
 
Location: Jerusalem

Join Date: Oct 2012
Posts: 1
Default library frgments size

Hi,
I'm preparing a library using NEB ChIP-seq kit. I'm starting with ssDNA of 250-800nt, making it double stranded using Klenow and random primers and then using the kit. The size of the fragments that I get after the PCR amplification is around 400bp (that includes the 120bp addition of the adapters and primers). I tried to start the library preparation with dsDNA the same size (250-800bp) and got the same 400bp library, so I don't think that the Klenow step is the problem.
Do you have any ideas why do I have selection for the short fragments and how to overcome it (since it is very important to me that the final library will represents all lengths).
Thanks
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