Hello All,
I work in a core sequencing facility (we do not perform Library preps) and we are having a problem with a customer's samples. It's actually 2 problems as we are having an issue with cluster density as it relates to loading concentration (normally loading 5 pM works very well but in this case it only generates ~ 30,000 clusters per tile). We quantitate using Qubit fluorencense and the Experion to get our size range correct and both returned expected results. The other issue, and these may be related, is that the pass filter % is very low (ranging to 70% to as low as 25%) and we observed the run as it took place and did not notice an abundance of clusters that would indicate overloading.
Does anyone have any experience with either issue and any methodology that could be used to remedy these problems. The client is getting anxious about his samples but I am wary of wasting more flow cell lanes until we can adress this issue. The sample is a yeast with a fairly balanced GC content.
Brian
I work in a core sequencing facility (we do not perform Library preps) and we are having a problem with a customer's samples. It's actually 2 problems as we are having an issue with cluster density as it relates to loading concentration (normally loading 5 pM works very well but in this case it only generates ~ 30,000 clusters per tile). We quantitate using Qubit fluorencense and the Experion to get our size range correct and both returned expected results. The other issue, and these may be related, is that the pass filter % is very low (ranging to 70% to as low as 25%) and we observed the run as it took place and did not notice an abundance of clusters that would indicate overloading.
Does anyone have any experience with either issue and any methodology that could be used to remedy these problems. The client is getting anxious about his samples but I am wary of wasting more flow cell lanes until we can adress this issue. The sample is a yeast with a fairly balanced GC content.
Brian
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