Mix libraries in a lane

[cascoamarillo]

Hi everyone,

I was wondering in someone has mix different library sizes in a single lane; and if so, is there any bias on the cluster formation?

What I'm thinking is to construct a small RNA library (20-35 nt) and a standard RNA library (around 100 nt); mix them together (of course the small RNA library should be made with adapters and primers so it can be sequenced with RD1 SP and index SP, and the same for the RNA seq experiment) and use a 60 cycles in the Illumina run.

Any thoughts?

Thanks

[pmiguel]

Not recommended. The cluster amplification process will almost certainly favor the shorter amplicons. They will produce very intense signal compared to the long amplicons. Likely this will cause issues.

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Phillip