Has anyone been able to successfully amplify their genomic libraries in 12 cycles or less using the Illumina or NEB reagents for library prep and show successful sequencing?
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Have you seen this paper? With modifications to the ligated adapters you can avoid PCR completely.
Amplification artifacts introduced during library preparation for the Illumina Genome Analyzer increase the likelihood that an appreciable proportion of these sequences will be duplicates and cause an uneven distribution of read coverage across the targeted sequencing regions. As a consequence, thes …
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I am wondering why they 5´-phosphorylate both adapters in that paper.
The adapter they call A_adapter_b needs it for ligation to the DNA fragment, but the other one ?
Am I missing something for which it might be required ?
seqgirl, I used 10 cycles with 5 ug starting material for paired-end genomic DNA sequencing - no problem.
However, I want to give the PCR-free a try.
Thank you in advance if someone has a hint !
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