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Old 08-03-2015, 08:53 AM   #1
Phuray
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Default HELP!!!! How stable are Nextera Index primers?

The problem has been solved, thanks a lot!

Last edited by Phuray; 08-17-2015 at 05:41 AM.
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Old 08-03-2015, 12:08 PM   #2
pmiguel
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They are probably fine, but if you don't want to take a chance, you could phone Illumina tech support and ask them. They might be willing to replace the primers.

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Old 08-10-2015, 07:13 PM   #3
GA-J
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I would simply test a few primers with index-PCR, and run on BA side by side.
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Old 08-13-2015, 11:36 PM   #4
Simone78
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they are quite robust, I have been freezing and thawing them probably 10 times or more and they were still fine. Unless you are using 50 ng of input DNA for your Nextera library prep, you have a huge excess of index primers anyway, so if part of them went bad, you are still safe.
We use the Nextera XT kit, start from <500 pg for tagmentation and dilute the primers 1:5 to ame them last longer. And, even then, if we donīt do a bead clean up afterwards, we still see a huge peak of unused adaptors in the prep.
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