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Old 04-29-2014, 07:04 PM   #1
Gina_P
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Default Illumina Nextera Mate Pair libraries.

I am curious about the directions for the Zymo Genomic DNA Clean & Concentrator recommended by the Illumina kit. It says to add 2 volumes of the DNA Binding Buffer to the tagmentation reaction and "mix thoroughly." What sort of mixing? flicking the tube? Pipetting? I am assuming that flicking the tubes is the safest thing to do here since I don't want to shear the DNA at this point.

Many thanks to anyone with experience and advice.

Gina
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Old 04-30-2014, 05:35 AM   #2
lorendarith
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Hi Gina,

Yes, you can try to flick the tube 5 times or so. Generally, flicking is not advisable when handling long DNA fragments, but they advise it throughout the whole Nextera MP prep.

If you really want to be sure your DNA gets minimal shearing/fragmenting while mixing - mix the liquid by pipetting with wide orifice tips. IMHO, this is the safest way.
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