Sigma Aldrich
DNase I
11284932001
Info: Browse DNase I Enzymes View Product
I have used DNase I from Sigma Aldrich to remove contaminating genomic DNA during total RNA extraction from the gram negative bacterial samples. After using DNase I at 10mg/ml concentration for 30 minutes during extraction step using Monarch Total RNA miniprep kit, genomic DNA was efficiently removed, which was checked by PCR using primers specific to a gene from gram negative bacteria (S. Typhimurium).
Removal of contaminating genomic DNA during total RNA extraction
10^10 S. Typhimurium cell pellets per column of Monarch total RNA miniprep kit
The protocol is very simple. After following the RNA extraction protocol steps from Monarch total RNA miniprep kit, add DNase I at 10mg/ml to the samples. Incubate for 30 minutes at room temperature and then follow the elution step to elute high quality genomic DNA free total RNA . The quality was assessed by PCR of gene specific to S. Typhimurium and agarose gel analysis revealed no amplification of the PCR product confirming efficient removal of genomic DNA.
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Efficient removal of genomic DNA contamination from total RNA samples as evidenced by PCR gel analysis of S. Typhimurium gene specific PCR. There was no amplification product and confirming efficient removal of genomic DNA.
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Efficient removal of genomic DNA contamination.
Overall, satisfied using DNase I from Sigma Aldrich to remove genomic DNA contamination from total RNA samples.