I have used a column based kit for isolating the RNA from the insect and plant sample. I got a good quantity of RNA, but in the sample QC for Library preparation, the sample failed due to poor RIN value. The Kit I used is specifically for the isolation of RNA of the highest integrity (RIN > 9) still it got a low RIN value.
Sample QC Details (Tape Station)
Sample - insect Qiaxpert (ng/μl) - 1162.9 A260/280 RATIO - 1.99 QUBIT (ng/μL) - 872 RIN Score - 3.9 COMMENTS (QC PASS/FAIL) - FAIL
Observation - Sample concentration outside the functional range for RINe and the assay; The upper ribosomal fragment has degraded.
Hi Vishali
if not degraded at start, salts, chemicals, enzymes, temperature, time...could have an impact. the best way is to make extraction over time in same conditions to allow standardization and comparisons between projects.
all the best
fred
Thank you, Frederic Lepretre. I am currently working on protocol standardization.
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