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Old 02-23-2011, 08:05 AM   #1
Location: New York

Join Date: Dec 2010
Posts: 40
Default Annealing index and primers for multiplexing??

We are planning to do a multiplexed cDNA library to run on Illumina . We bought the indexes and primers from IDT (sequences from Illumina) but we have some questions. We were reading that Illumina changed the protocol and now it uses the index+PCRprimer (all together) instead of using the index as a "third" primer.
We were thinking about annealing the index+primer and use it as a single one but we are not sure if it is gonna work. Does anybody have tried that?
If so, which temperature?
Otherwise we will use as separated things. Has anybody got good results with this?

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Old 03-09-2011, 07:51 PM   #2
Location: London

Join Date: Sep 2010
Posts: 22

ahhh...we used to do this but not anymore. Just couldn't get good annealing, we'd see multiple peaks. I recommend using the TruSeq adapters from Illumina or the DNA Adapters from Bioo which are sold separately from their library prep kit.
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annealing, illumina, index, multiplexing, primer

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