Latest revision as of 17:40, 15 August 2013

Plasmid Isolation by ethanol precipitation

Follow QIAprep Spin Miniprep Kit protocol from steps 1 - 4.
Centrifuge for 1h at 4C and 20000 g.
Carefully transfer the supernatant to a new Eppendorf. Be careful not to take any solid parts (this is the cell debris).
To the sample add 3 volumes of ice-cold 70% ethanol. If needed divide into several samples.
Put samples to -20 for 0.5h to 1 h to precipitate DNA.
Centrifuge at 4C for 30 mins at 20000 g.
Remove the supernatant.
Add 300 uL of 90% ice-cold ethanol. Do not pipette up and down.
If the pellet is detached centrifuge for 5 to 10 mins at 4C and 20000 g.
Remove the ethanol with a pipette.
Let the sample stand open to dry for up to 5 mins.
Resuspend in 50 - 100 uL water.

@@ Line 1: / Line 1: @@
-{{:Team:DTU-Denmark/Templates/StartPage|Plasmid Isolation}}
+{{:Team:DTU-Denmark/Templates/StartPage|Plasmid Isolation by ethanol precipitation}}
+<div class="overviewPage">
+# Follow QIAprep Spin Miniprep Kit protocol from steps 1 - 4.
+# Centrifuge for 1h at 4C and 20000 g.
+# Carefully transfer the supernatant to a new Eppendorf. Be careful not to take any solid parts (this is the cell debris).
+# To the sample add 3 volumes of ice-cold 70% ethanol. If needed divide into several samples.
+# Put samples to -20 for 0.5h to 1 h to precipitate DNA.
+# Centrifuge at 4C for 30 mins at 20000 g.
+# Remove the supernatant.
+# Add 300 uL of 90% ice-cold ethanol. Do not pipette up and down.
+# If the pellet is detached centrifuge for 5 to 10 mins at 4C and 20000 g.
+# Remove the ethanol with a pipette.
+# Let the sample stand open to dry for up to 5 mins.
+# Resuspend in 50 - 100 uL water.
-# Follow QIAprep Spin Miniprep Kit protocol from steps 1 - 4
+<div style="clear: both;"></div>
-# Centrifuge for 1h at 4C and 20000 g
+</div>
 {{:Team:DTU-Denmark/Templates/EndPage}}