Trusight One panel Illumina

[María Camila Fetiva]

Hello, I have been working with trusight One panel from illumina, and it has been so difficult to standarize, and we have had oversultering and underclustering issues. We are working with a Miseq. We would like to be in contact with a person with experience in trusight One panel to have a scientific consultancy. We could pay for some courses or consultacy specificly about trusight one. Please let me know if some one of you has this experience. Thank you so much and could help us to solve this problematic

[finswimmer]

Hello,

how many samples have you pooled? In what concentration did you load your library? What was the resulting cluster density and cluster passing filter?

fin swimmer

[María Camila Fetiva]

Hello we have pooled 3 samples, we loaded library in 1.25nM (1 assay with overlustering CLusters Density: 1909 k/mm2, PF: 67%), after that we loaded library to 1.0nM (Clusters Density: 137 k/mm2, PF: 98%).

If you want i could send you by email a table with the resume of the failed Runs.

Thank you so much for your help.

[finswimmer]

Hello,

are you sure you loaded 1.25nM? Normal final concentrations of the library on a MiSeq are between 8-16pM.

What was the concentration of the library at the end of the library preparation? How did you quantify?

What is the average fragment size? How did you measure it? Whith Bioanalyzer/Tapestation? Can we see the profile?

fin swimmer