Roche Diagnostics, Germany
DNase 1 recombinant, RNase-free
04716728001
Aim of the study was to purify RNA from rat liver for gene expression studies. The RNA was isolated using TRI reagent. To the isolated RNA, DNase 1 recombinant, RNase-free solution was added to remove the contaminating DNA.
Preparation of purified RNA for gene expression analysis using real time PCR
Rat liver tissue, TRI reagent, 1-Bromo-3-chloropropane, isopropanol, Nuclease free water
RNA from rat liver was isolated using TRI reagent. 100 mg of tissue was homogenized 1 ml of TRI reagent. To the homogenate, 1 ml 1-Bromo-3-chloropropane was added, mixed vigorously and centrifuged. To the clear aqueous phase isopropanol was added to precipitate the RNA. The RNA pellet obtained was washed with 75% ethanol. Finally, the RNA pellet was air dried and dissolved in appropriate volume of sterile nuclease free water. To the RNA, DNase 1 recombinant, RNase-free was added according to the manufacturer’s instruction and incubated at 25 to 37 °C for 15-30 min and reaction was stopped by 0.2mM EDTA and DNase 1 recombinant was inactivated by 10 min incubation at 75 °C. RNA was quantified by nanodrop and the RNA integrity was checked by gel electrophoresis.
None
The DNA free RNA was isolated with purity (ʎmax 260/280) of nearly 1.8 and the integrity of isolated RNA was also maintained as checked by gel electrophoresis.
Helps to get DNA free RNA which is perfect for real-time PCR
No
DNase 1 recombinant, RNase-free from Roche Diagnostics is suitable for purifying RNA