Difference between revisions of "20.109(S11):Purify RNA and run affinity column (Day4)"
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#*If the buffer won't start flowing, talk to the teaching faculty. Sometimes pushing the cap on and then off will help it begin. | #*If the buffer won't start flowing, talk to the teaching faculty. Sometimes pushing the cap on and then off will help it begin. | ||
#When the buffer stop flowing, discard the eluant into a waste tube and centrifuge the tube for 2 min at 1000 g. (No need to replace the cap.) | #When the buffer stop flowing, discard the eluant into a waste tube and centrifuge the tube for 2 min at 1000 g. (No need to replace the cap.) | ||
− | #*''Is g the same as rcf or rpm? Ask if | + | #*''Is g the same as rcf or rpm? Ask if you re not sure!'' |
#Add 500 μL of Selection Buffer to the top of the column. Let it drain by gravity for a couple of minutes, then spin for 1 min at 1000 g. | #Add 500 μL of Selection Buffer to the top of the column. Let it drain by gravity for a couple of minutes, then spin for 1 min at 1000 g. | ||
#Repeat step 7 three more times. Meanwhile, label two 1.5 mL eppendorf tubes for collection (on the side ''and'' with a sticky label on the cap indicating the RNA sequence identity), and cut off their caps. | #Repeat step 7 three more times. Meanwhile, label two 1.5 mL eppendorf tubes for collection (on the side ''and'' with a sticky label on the cap indicating the RNA sequence identity), and cut off their caps. | ||
− | #Transfer each column to a 1.5 mL tube | + | #Transfer each column to a 1.5 mL tube |