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Old 05-09-2017, 05:21 AM   #1
christophpale
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Location: canada

Join Date: May 2010
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Default Library preparation for Amplicon sequencing by HiSeq

Hello,
I have 60 amplicons of around 300bp that I need to perform deep sequencing to find mutations.
Can I just synthesize the P7 and P5 adaptors from IDT, ligate onto my amplicons and send out for sequencing? Or do I need to actually buy the Truseq custom amplicon kit (https://support.illumina.com/sequenc...amplicon.html?

Thank you
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Old 05-09-2017, 04:58 PM   #2
nucacidhunter
Jafar Jabbari
 
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If you already have generated your amplicon pool, you just need to prepare a library using any shotgun DNA library prep kit skipping size-selection step. For best results amplicons need to go through end-repair and A tailing steps before ligating adapters.

Illumina's custom amplicos is suitable when there are many samples and large target region.
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