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Old 07-06-2011, 06:22 AM   #1
klytie
Junior Member
 
Location: Tallinn

Join Date: Jul 2011
Posts: 5
Default T4 kinase in end repair protocol

Hi!
I'm new in this, so forgive me if my question is stupid. Why do I need to add T4 PNK to the mixture of Klenow and T4 polymerase when repairing the ends of enzyme fragmented library. I'm trying to put together a protocol for Illumina sequencing but at the moment we are not using NEB next or Illumina or any kind of kit prepared for end repairing. There are only separately bought enzymes on the menu. If I remember correctly, I have seen protocols with and without T4 PNK. When do I need to use this enzyme and when is it not necessary to put thisone in? Can anyone also provide me the units per end repair reaction for T4 pol and Klenow?

Many thanks!
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