We ran unrelated libraries in the individual quadrants on one plate. I see few sequences clearly belonging to a library from other quadrants. I just want to make sure if the only explanation is contamination. It is clear to me that it is the most obvious.
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I always wondered if the gasket was able to completely isolate each region of the plate. For well over a year, the 2-lane gasket would move in every centrifugation. We tried all sort of things, including using different BDDs. The problem was only solved after we stopped sealing the ports.
What was the percentage of contamination? -
My quadrant was number 4. Number three has about 1% contamination of my sequences (3K out of 300K). The quadrants 1 and 2 have lower contamination, few hundred sequences.
I do not have the details of the plate arrangement and handling, the run was done by the university core facility. They are generally very carefull and I do not think they would contaminate during library prep.Comment
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