Real Time PCR From Cell Culture: Difference between revisions
→References (Saltiel Lab): added Chang et al Mol Endocrinol |
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==Protocol== | ==Protocol== | ||
===RNA Extraction=== | ===RNA Extraction=== | ||
#Use RNEasy kit with Qiashredder. For a 12 well plate, use 350 uL of RLT (add 10 uL B-ME per 1 mL of RLT buffer). | #Use RNEasy kit with Qiashredder. see [[Harvesting RNA from Cells grown in monolayer]]. For a 12 well plate, use 350 uL of RLT (add 10 uL B-ME per 1 mL of RLT buffer). | ||
#Scrape cells and pass through Qiashredder column. Do the optional DNAse step at step 5 using the RNAse free DNAse from Qiagen. | #Scrape cells and pass through Qiashredder column. Do the optional DNAse step at step 5 using the RNAse free DNAse from Qiagen. | ||
#Store at -20 until RT reaction | #Store at -20 until RT reaction | ||