Sigma
TRI Reagent
T9424
MCF7 cells were grown in 6 well plate. Once it reaches confluence, cells were washed with PBS and total RNA was isolated by TRI Reagent.
Total RNA isolation
MCF7 cells
MCF7 cells were grown in 6 well plate. Once it reaches confluence, cells were washed with PBS and 1 ml of tri reagent was directly added to each well and cells were lysed. The lysate was collected in 1.5 ml centrifuge tube. 0.2 ml of Chloroform wad added to each tube and mixed and centrifuged. The mixture separates into 3 phases; an aqueous phase containing RNA, the interphase containing DNA and an organic phase containing protein. The aquesous phase collected and isopropanol was added to precipitate RNA.
RNase free environment is essential - clean the working area with DEPC
Good quality total RNA was isolated which was confirmed by A260/A280 ratio.
N/A
Best reagent for total RNA isolation
Need to optimize proper input amount
Tri reagent is best for high quality total RNA isolation