Qiagen
RNAprotect Bacteria Reagent
76506
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I have used Qiagen RNAprotect Bacteria Reagent for stabilization of total RNA prior to RNA extraction for qPCR experiments. At the end of extraction, the quality of RNA was assessed by Agilent Bio-analyzer as well as qPCR for selected gene expression. Gene expression was completely variable (not constant levels in normalizer gene) in case of samples without RNA protect treatment.
RNA extraction for qPCR
E. coli culture grown in LB broth to mid-log phase
3 ml of E. coli culture grown in LB broth were mixed with or without 6 ml of RNA protect reagent (1:2 ratio for culture: RNA protect) and then pelleted at 4000 rpm at 4 degrees Celsius. These pellets were then used for RNA extraction using RNeasy Midi kit according to kit's instructions. Finally eluted in 200 microliter of nuclease free water.
After adding RNA protect to culture, mix well and allow it to stand at room temperature for 30 minutes.
Stabilization of RNA was found by performing qPCR on cDNA prepped from RNA extracted from RNA protect used group and without the RNA use of RNA protect. There was wavy gene expression for normalizer gene in the case cDNA prepped from RNA samples without RNA protect. Agilent Bio-analyzer showed RIN values of 8 to 9.2 for those extracted with RNA protect.
None
Very easy to use. Simple method.
RNAprotect Bacteria Reagent is a must for RNA extraction from bacterial samples.
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