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Old 07-17-2013, 07:24 AM   #8
HelenaSC
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Location: Spain

Join Date: Jun 2013
Posts: 21
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My last question before starting (sorry!!! ):

I have two samples (1.1nM) that I want to run together on the MiSeq. Following your instructions, do you think this will be ok ?(to end up with a 11pM final concentration):

pool 2 libraries (1.1nM each one) :

5ul lib1 + 5 ul lib2 = 10ul pool 1.1nM
+ 2ul 0.5N NaOH
1ul water
(13ul of denatured samples at 0.84nM)

Then, neutralize with 1ul of 1N HCl (denatured and neutralized samples ; 14ul at 0.78nM)
Finally, dilute samples with 992ul HT1 buffer = samples at final concentration= 11pM (load 600ul).

Thanks a lot!!!
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