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Old 07-19-2012, 01:13 PM   #1
lee27
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Location: Ithaca, NY

Join Date: Jul 2012
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Default Ligating barcodes to amplicons

Hi all,
I am new to the forum and to next-gen in general. I am working on preparing a library of amplicons (20 different amplicons per individual sample - 96 to 180 samples) and I want to barcode the amplicons so that I can identify which individual they are from. Can barcodes/indices be directly ligated to the poly-A tail resulting from PCR or do I first need to blunt the end, apply a poly-A tail and then ligate? Thanks for your thoughts and patience!
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Old 07-24-2012, 10:43 AM   #2
Zaag
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Location: Amsterdam

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Yes I would alwasy make it blunt end and perform the A-tailing. You could also design fusion primers with the adapter and MID so after PCR you can start emPCR without ligating.
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