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Old 04-08-2016, 10:46 AM   #13
lac302
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Location: DE

Join Date: Dec 2012
Posts: 65
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Quote:
Originally Posted by nucacidhunter View Post
I think the only difference is concentration of enzymes. Following to consider:
1- The buffer used to elute from columns. If TE has been used, inhibition of tagmentation enzymes is expected.
2- If you have enough DNA you can do another clean up with 1.2x AMPure bead or a column and elute in TE0.1
3- GC content of genome, normally %GC 30-75 is OK
I think Qiagen EB buffer was used. Either way these samples are fairly concentrated. I end up diluting close to 500x in water. GC content is around 48%, so no issues there.

Quote:
Originally Posted by sweetph3 View Post
Any time I've had issues, especially if the gDNA was prepared by another group, I start off with the MoBio Power Clean now. It hasn't failed me yet. I think some samples have inhibitors that carry over even through columns. Good luck!
It's definitely an inhibitor issue. Possibly carbohydrates...We have powersoil kits on hand that I'm trying today.
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