Balancing run with nanoflow cell

[ramarquezo]

Hi everybody, I'm new in Illumina MiSeq, I have one question, what is the meaning of to do a previous balancing of the run of a multiplexing library by mean of nanoflow cells. In a process in my institution will be used a previous step of balancing using a nanoflow cell, and after this will be done the sequencing process using a full flow cell run. I don't understand why the previews process using the nanoflow cell could balancing the run?

[kcchan]

I guess what your institution is doing is using a short MiSeq run to check the evenness of multiplexed pools. So for example if there is a 10 sample pool they would be looking to see of each sample gets about 10% of the total number of reads. The data would also be useful to look at other things like insert sizes and screening for contaminants like rRNA in RNA-Seq libraries. While this may be helpful to troubleshoot a bad library, it's usefulness diminishes as the level of multiplexing goes up since it would be very difficult to re-adjust any one particular sample with precision.

[ramarquezo]

Thanks

Thanks Kcchan, your answer is very useful, is so much coherent!!!

[koadman]

Hi ramarquezo, kcchan, yes that is indeed what we typically do here at UTS. Prep lots of libraries, QC them by generating a little bit of data on a nano flowcell, then balance them precisely for a subsequent full flowcell run. ramarquezo, feel free to swing by my office and introduce yourself sometime if you have more questions!

[ramarquezo]

Thanks Koadman! Your answer give me a total clarity of the process that we are doing!!