Skew ratio

[neelam bodhale]

Hello everyone,

First of all, I am very much blessed to be part of this forum. I came across this forum while I was troubleshooting an issue related to PCR of GeCKO v2. 

I've recently begun working with the GeCKO v2 mouse gene knockout library. Prior to my involvement, a colleague amplified the library twice, but the skew ratio between the top 10% and bottom 10% was quite high. I've attached the analysis results and would appreciate any expert insights on these analysis results. I would like to know which areas need to be improved and if there is a way to improve that.

I have since re-amplified the library, achieving high transformation efficiency. We are currently awaiting the sequencing results.

Looking forward to suggestions.

Thank you.

Best regards.

Neelam

[neelam bodhale]

Hi all,

Here is another set of sequencing data of different batches of amplified GeCKO v2 mouse library.

Thank you 

Neelam