Qiagen
RNeasy Midikit (50)
75144
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I used Qiagen's RNeasy Midikit for extraction of total RNA from the Salmonella enterica cultures grown in LB medium with different levels of bile salts in the medium. This kit method gave very high quality RNA (RIN numbers 8.2 to 9.2) and good yield (85 microgram total in 200 microliter elution).
Extraction of Total RNA from Salmonella enterica cultures for qPCR experiments
3 ml of Salmonella enterica culture grown in LB broth stabilised with 6 ml of RNA protect bacteria reagent
Bacterial culture stabilised with RNA protect bacteria reagent was centrifuged at 4000 rpm for 10 minutes at 4 degree C. Supernatent after centrifugation was removed and pellet lysed according to kit's instructions given. Then followed lysis-on column DNA digestion, washing and elution steps using appropriate reagent solutions given in the kit. Finally total RNA from midikit columns was eluted in 200 microliter of RNase free water. Then quality and quantity of eluted RNA was assessed by Agilent Bioanalyzer analysis and Nanodrop analysis.
Perform on-column DNA digestion using DNase enzyme to remove genomic DNA contamination during RNA extraction
Got very good quality total RNA. RIN values of 8.2 to 9.2 were noticed for all the samples as assessed Agilent Bioanalyzer analysis. Yield was 85 microgram total in a 200 microliter volume. Nanodrop 260:280 ratio was 2.24.
None
Entire protocol is very easy to follow and Perform.
Very good Qiagen kit for very good quality extraction of RNA from the bacterial cultures.
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