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Old 07-21-2016, 08:18 AM   #15
cerbatana
Junior Member
 
Location: Tokyo

Join Date: Jun 2016
Posts: 6
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EDIT:

I just realized that the original post was about Nextera kit while the one I' using is Nextera mate-pair kit. I'll create a new thread so this post can be removed. Thank you.

Hi! I'm also having problems with the same library preparation kit and I really don't know if the enzyme is working at all. I' attaching the bioanalyzer DNA 12000 results right after the tagment genomic DNA step. I tested two different lots of the enzyme but the results are the same, and I also don't think is a DNA quality problem because this DNA has been successfully used with the Nextera XT kit which also uses a tagmentation step.

I used 4 ug of DNA as input (good quality according to PFGE results with fragments longer than 50 kbp) and 4 uL (lane 1) or 8 uL (lane 2) of tagment enzyme from one lot and 4 uL from another lot (lane 3). Lane 4 was loaded with water.

There seems to be something in the 17000 bp region but I don't know if this long fragmented DNA or unfragmented DNA that migrated with the longer band of the ladder.

What happened?
Attached Images
File Type: png tagment.png (137.6 KB, 55 views)

Last edited by cerbatana; 07-21-2016 at 08:45 AM.
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