Hi Everyone I am newbie here and glad I found this wonderful and informative forum. I was wondering if some of you guys can help me out with my MiSeq run stats?
I am using the MiSeq v2 300cycle kits with a targeted sequencing pool of 5 libraries (generated by the Fluidigm Access Array system). I have diluted the pool to 8pM (apparently this gives optimum cluster density with fluidigm libs) and spiked in 5% 12.5pM PhiX. I am also using Fluidigms custom primers.
It is worth mentioning that the kits are out of date by 2 months which may be causing the errors (they were provided to us for free while we are awaiting for new kits to be delivered).
I put the run on 24hrs ago so it is nearing completion and I am getting the following stats:
Cluster density: 988+/-235, Cluster PF: 6.31+/-13.08, Phas/Prephas: 0.288/0.087, Reads: 21.56, Reads PF: 0.93, Q30: 90.8%
The cluster density seems to be just within the optimum range and the Q30 is quite nice but the PF is terrible and I am unsure of why this is the case? Could anyone help me with possible reasons?
I am using the MiSeq v2 300cycle kits with a targeted sequencing pool of 5 libraries (generated by the Fluidigm Access Array system). I have diluted the pool to 8pM (apparently this gives optimum cluster density with fluidigm libs) and spiked in 5% 12.5pM PhiX. I am also using Fluidigms custom primers.
It is worth mentioning that the kits are out of date by 2 months which may be causing the errors (they were provided to us for free while we are awaiting for new kits to be delivered).
I put the run on 24hrs ago so it is nearing completion and I am getting the following stats:
Cluster density: 988+/-235, Cluster PF: 6.31+/-13.08, Phas/Prephas: 0.288/0.087, Reads: 21.56, Reads PF: 0.93, Q30: 90.8%
The cluster density seems to be just within the optimum range and the Q30 is quite nice but the PF is terrible and I am unsure of why this is the case? Could anyone help me with possible reasons?
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