View Single Post
Old 04-16-2014, 03:34 PM   #1
arilindsey
Junior Member
 
Location: California

Join Date: Apr 2014
Posts: 1
Default Nextera Kit -- DNA fragments too small

Hello --

I have been working with the Nextera kit for Illumina to create DNA libraries for genome sequencing. I initially used the recommended 50ng input DNA (quantified with a Qubit). However, what should have been my final library had no DNA, and instead all the DNA was left in the supernatant that comes off the first Ampure XP bead clean up step. When I run that same supernatant on a gel, the DNA fragments range from 80 - 200 bp. I have tried higher concentrations of input DNA (up to 90ng) and still have the same problem. Not sure what to try next -- has anyone experienced this? My input DNA has a high molecular weight -- 20kb+ fragments.

Any help is much appreciated.
Cheers,
Amelia
arilindsey is offline   Reply With Quote