Difference between revisions of "Ethanol Precipitation of Vector DNA"

Latest revision as of 14:53, 4 June 2015

After digestion with restriction enzymes (ex. Normally a 40 µL digestion)
Increase volume to 90 µL with dH20
Add 10 µL of 3M NaOAc pH 5.2
Add 2X the volume of ethanol (ex. 200 µL of ethanol)
Vortex and put in -80°C freezer for 15 minutes
Centrifuge for 30 minutes (Place the tube hinge out in the microcentrifuge to know where the pellet is)
Dump off the liquid and allow to dry, either sitting on the bench or with the lyophilizer

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-•	After digestion with restriction enzymes (ex. Normally a 40 µL digestion)
+# After digestion with restriction enzymes (ex. Normally a 40 µL digestion)
-•	1/10 the digestion volume of NaOAc (ex. 4 µL of NaOAc)
+# Increase volume to 90 µL with dH20
-•	2X the reaction volume of ethanol (ex. 88 µL of ethanol)
+# Add 10 µL of 3M NaOAc pH 5.2
-•	Vortex and put in -80°C freezer for 15 minutes
+# Add 2X the volume of ethanol (ex. 200 µL of ethanol)
-•	Centrifuge for 15 minutes (Place the tube hinge out in the microcentrifuge to know where the pellet is.
+# Vortex and put in -80°C freezer for 15 minutes
-•	Dump off the liquid and allow to dry, either sitting on the bench or with the lyophilizer.
+# Centrifuge for 30 minutes (Place the tube hinge out in the microcentrifuge to know where the pellet is)
+# Dump off the liquid and allow to dry, either sitting on the bench or with the lyophilizer