We did not get the RNA pellet after the fase separation. Seems Very difficult to lyse the cells.
Vidyanand Jha
Harvesting Cells:Grow Geobacillus/Parageobacillus cultures to the desired OD600. Pellet cells by centrifugation at a suitable speed and duration. Cell Lysis:Resuspend the cell pellet in Trizol reagent, following the manufacturer's instructions. Add chloroform and perform phase separation. RNA Precipitation:Transfer the aqueous phase to a new tube and precipitate RNA with isopropanol. Centrifuge and wash the RNA pellet with 75% ethanol. RNA Purification:Air-dry the RNA pellet or use a speed vacuum. Dissolve RNA in RNase-free water or an appropriate buffer. DNase Treatment:Treat RNA with DNase I to remove genomic DNA contamination. RNA Quality Control:Assess RNA integrity and purity using gel electrophoresis or a bioanalyzer. Quantification:Quantify RNA concentration using a spectrophotometer or fluorometer.
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