View Single Post
Old 04-08-2016, 10:46 AM   #13
Location: DE

Join Date: Dec 2012
Posts: 65

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.

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.
lac302 is offline   Reply With Quote