Real Time PCR From Cell Culture: Difference between revisions
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===Plate Preparation=== | ===Plate Preparation=== | ||
#Book 3h on qPCR machine at http://felix2.lsi.umich.edu/ORS | #Book 3h on qPCR machine at http://felix2.lsi.umich.edu/ORS | ||
#Prepare primer mix from 5 uM primer pair stocks | #Prepare primer mix from 5 uM primer pair stocks. Per reaction add 0.4 uL primers + 4.6 uL water. | ||
#Get 96 well block and keep on rack. Do not touch bottom of plate. | #Get 96 well block and keep on rack. Do not touch bottom of plate. | ||
#Add 5 uL template per well. | #Add 5 uL template per well. | ||
#Add 5 uL primer per well | #Add 5 uL primer per well. | ||
#Add 10 uL per row into 8 wells of a PCR strip plus an extra 10-20 uL | #Add 10 uL per row into 8 wells of a PCR strip plus an extra 10-20 uL | ||
#Using a multichannel pipettor, add 10 uL Master mix to each well | #Using a multichannel pipettor, add 10 uL Master mix to each well | ||