Hello everyone.
Lurked for a while, but first time posting. I have a bit of a vexing issue.
We are using a MiSeq Micro kit (v2 chemistry, 300 cycles).
Libraries were mixed, denatured, PhiX added, loaded on the cartridge, etc.
(15pM loading, 5% PhiX @ 10pM)
The run started and it completed, but did not generate any data except cluster density (cluster density around 900, which I have already determined why it might be low).
-no cluster diversity
-no tile images
-no % PF
-no % aligned
-no sequence data
-no undetermined bins
I don't believe it is a sample issue as the libraries were quantified using kapa, looked at via Tapestation, and re-quantified by Qbit for good measure (just to get library quality out of the way as a reason). and adjusting for a slight mistake calculating the library concentrations, the cluster density is what I would expect had the rest of the run work.
PhiX had been previously denatured and used before and after this run with success.
We have never used the Micro kit before and tech support had suggested there was nothing special you had to do or change in the sample sheet or LRM (since the kit's RFID code is read by the machine), but it is the only logical place that makes sense. This is based on the fact that the PhiX had worked on other runs that were using the Micro kit without issues, and for the % aligned to not show up with the PhiX makes me think there was a setting or something that made it so no data was collected.
Has anyone ever seen an issue like this before? Could really use some help as I have been beating my head against my desk for the last 2 weeks.
Lurked for a while, but first time posting. I have a bit of a vexing issue.
We are using a MiSeq Micro kit (v2 chemistry, 300 cycles).
Libraries were mixed, denatured, PhiX added, loaded on the cartridge, etc.
(15pM loading, 5% PhiX @ 10pM)
The run started and it completed, but did not generate any data except cluster density (cluster density around 900, which I have already determined why it might be low).
-no cluster diversity
-no tile images
-no % PF
-no % aligned
-no sequence data
-no undetermined bins
I don't believe it is a sample issue as the libraries were quantified using kapa, looked at via Tapestation, and re-quantified by Qbit for good measure (just to get library quality out of the way as a reason). and adjusting for a slight mistake calculating the library concentrations, the cluster density is what I would expect had the rest of the run work.
PhiX had been previously denatured and used before and after this run with success.
We have never used the Micro kit before and tech support had suggested there was nothing special you had to do or change in the sample sheet or LRM (since the kit's RFID code is read by the machine), but it is the only logical place that makes sense. This is based on the fact that the PhiX had worked on other runs that were using the Micro kit without issues, and for the % aligned to not show up with the PhiX makes me think there was a setting or something that made it so no data was collected.
Has anyone ever seen an issue like this before? Could really use some help as I have been beating my head against my desk for the last 2 weeks.
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