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Hi,
We're doing a quick & dirty transcriptome for a low-complexity tissue using a single library on a MiSeq; time/cost are a factor. The run is a single sample RNA-Seq; V2 reagent; 2x150 PE. Illumina recommends 12.5 pM for their PhiX control, but in the interest of not overloading the run, I'm thinking of about a 10 pM load with a 2% PhiX (@ 10 pM stock) spike-in. Would I be pushing the upper limit - the library fragment size (by KAKA) is 470 bp. I don't normally do a transcriptomes/RNA-Seq on the MiSeq, and thus the question. Thoughts?
Thanks...
We're doing a quick & dirty transcriptome for a low-complexity tissue using a single library on a MiSeq; time/cost are a factor. The run is a single sample RNA-Seq; V2 reagent; 2x150 PE. Illumina recommends 12.5 pM for their PhiX control, but in the interest of not overloading the run, I'm thinking of about a 10 pM load with a 2% PhiX (@ 10 pM stock) spike-in. Would I be pushing the upper limit - the library fragment size (by KAKA) is 470 bp. I don't normally do a transcriptomes/RNA-Seq on the MiSeq, and thus the question. Thoughts?
Thanks...