QPCR: Difference between revisions
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#You can prepare the plate ~3h beforehand, keeping it at 4C | #You can prepare the plate ~3h beforehand, keeping it at 4C | ||
#Immediately before the run spin the plate briefly in the swinging bucket centrifuge. | #Immediately before the run spin the plate briefly in the swinging bucket centrifuge. | ||
===Run Protocol=== | |||
#Open LightCycler software. | |||
#Click new experiment. | |||
#Choose SYBR Green option from drop down and make sure that the correct block is in place (it will say 96 or 384). | |||
#Click apply template. | |||
#Go to templates>run templates> bridges SYBR Green 384. | |||
#Load plate and click 'start run'. | |||
#This protocol is set to do the following: | |||
#Activate- 1 cycle @95 degrees for 10s. | |||
#Amplify- 45 cycles @95, 60 and 73 degrees for 15s, 15s and 10s, respectively. | |||
#Melt- 1 cycle @95, 40, 65, 95 and 40 degrees for 1m, 1m, 1s, continuous and 10s, respectively. | |||
==Calculations== | ==Calculations== | ||