Hi everyone,
I have a question regarding the pooling of NexteraXT and TrueSeq PCR free libraries onto a single Illumina HighSeq lane. I am sharing a lane with a colleague and we have different needs: I have enough DNA so I would like to use PCR free libraries. My colleague on the other hand has very little DNA and wants to use NexteraXT preps. Each of use would like to use 1/2 of a lane for sequencing.
The person at our sequencing facility told us that pooling PCRed and PCR-free libraries could be problematic because it is hard to balance molarities in such a way that the different library types each occupy 1/2 of a lane.
Now we are wondering about the possible reasons behind this? Does anyone have an idea why this could happen. I have never seen this mentioned in a paper.
To which extent is this really a problem? When aiming for a 50:50 lane split how far off could the output be? 60:40? 80:20?
thank you all for your help!
I have a question regarding the pooling of NexteraXT and TrueSeq PCR free libraries onto a single Illumina HighSeq lane. I am sharing a lane with a colleague and we have different needs: I have enough DNA so I would like to use PCR free libraries. My colleague on the other hand has very little DNA and wants to use NexteraXT preps. Each of use would like to use 1/2 of a lane for sequencing.
The person at our sequencing facility told us that pooling PCRed and PCR-free libraries could be problematic because it is hard to balance molarities in such a way that the different library types each occupy 1/2 of a lane.
Now we are wondering about the possible reasons behind this? Does anyone have an idea why this could happen. I have never seen this mentioned in a paper.
To which extent is this really a problem? When aiming for a 50:50 lane split how far off could the output be? 60:40? 80:20?
thank you all for your help!
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