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4 years ago
4 years ago by Paul G. Macon
Hello! I used Trizol to extract total RNA from in-vitro cultured bacteria (1 X 10^8 cells). After phase separation, I mixed ~0.4 ml of the upper phase which contains RNA with 0.5 mL cold isopropanol. However, the amount of RNA when measured in Nanodrop was very low. In addition, the ratio between 260 and 230 was around 0.1 to 0.5. Is there a chance that my sample was contaminated by the Trizol reagent? When I collected the aqueous phase I made sure to not touch the lower phase. What should I do?
Found 4 matching solutions for this experiment
|For retrieval of RNA from the extraction solvent following mechanical disruption, we compared simple isopropanol precipitation of RNA to solid-phase isolation of RNA using the Purelink miRNA Isolation Kit (Invitrogen, Carlsbad, CA, USA). A summary of all conditions tested and their respective nucleic acid yields before DNase I treatment as assessed by UV spectroscopy is detailed in Supplementary Table S1; the analyses were performed in three biological replicates.|
|To yield high RNA, give the column an extra wash with both RW1 and RPE buffers (3 total washes with each buffer).|
|"Include DNAse treatment for 15-20min.
Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C.
Use water to elute the RNA that is warmed to ~60`C"
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