View Single Post
Old 06-30-2016, 02:44 PM   #4
nucacidhunter
Jafar Jabbari
 
Location: Melbourne

Join Date: Jan 2013
Posts: 1,238
Default

Quote:
Originally Posted by microgirl123 View Post
I don't get the same results from the BR and HS assays either. I think the HS assay is just not correct at the top end of the standard curve. I usually pool samples based on the HS assay and then use qPCR to quantify the pooled library - my pooled library is often twice as concentrated as it should be based on the Qubit.
Qubit and qPCR quantify different properties of library. Qubit quantifies dsDNA regardless of adapter presence in fragment termini while qPCR quantifies the fragments flanked by adapters that are amplifiable. The quantity measured by these methods will be closer to each other if all fragments were double stranded and flanked by adapter sequences.

Qubit will be fine for amplicon libraries quantification where the PCR input was low so there is not much carry over of non-adapted DNA.

Last edited by nucacidhunter; 06-30-2016 at 03:03 PM.
nucacidhunter is offline   Reply With Quote