Hey I was wondering if anyone has any ideas as to why I ended up with very small quantities of finished library after starting with 1 ug total RNA per sample and after following the low sample protocol in this protocol https://www.utsouthwestern.edu/labs/next-generation-sequencing-core/assets/truseq-stranded-mrna-sample-prep-guide.pdf
Library concentrations/amounts that I had:
- Concentration (nanograms/microliter) Amount (in nanograms)
- 0.924 ng/ul 17 ul * 0.924 ng/ul = 15.708 ng
- 1.08 ng/ul 17 ul * 1.08 ng/ul = 18.36 ng
- 3.40 ng/ul 29 ul * 3.40 ng/ul = 98.6 ng
- 1.16 ng/ul 29 ul * 1.16 ng/ul = 33.64 ng
- 0.284 ng/ul 29 ul * 0.284 ng/ul = 8.236 ng
Also had primer dimer peaks when I ran an aliquot of each library on a bioanalyzer 2100 platform that did not go away when doing a second bead clean up with the AMPure XP beads for the first two samples
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