Revision as of 22:37, 28 September 2011

Plasmid Extraction

This protocol is utilized in accordance to the manufacturer's protocol from Omega E.Z.N.A (EaZy Nucleic Acid Isolation)

Place gel on the UV box
Carefully extract the fragment suspended in the gel
Mass gel fragments
Place fragment into a 1.5 mL tube and add 4 µL of H2O
Volume of water added to volume of gel is 200% however if fragment it small 1 mL of water will suffice
Remove H2O
Add equal amounts of H2O and Binding Buffer (XP2) to the gel
Incubate mixture at 55 degrees for 7 mins
Mix with vortex for 2 mins
Place in the HiBind DNA Mini Column in the 2 mL tube
Add 700 µL at 10,000xg for 1 min
Discard liquid
Add 300 µL Binding Buffer (XP2) into the HiBind DNA Mini Column and spin down at 10,000xg for 1 min
Discard liquid
Wash the column with 700 µL of SPW buffer with added ethanol and spin down at 10,000xg for 1 min
Discard liquid
Wash the column with 700 µL of SPW buffer again and spin down at 10,000xg for 1 min
Discard the liquid
Spin down the column at 13,000xg for 1 min to dry the column
Elute in 50 µL of H2O and wait 1 min
Spin down the column at 13,000xg for 1 min to dry the column
Use a spectrophotometer to measure the concentration and the purity of your plasmid

@@ Line 12: / Line 12: @@
 <ol>
 	<li> Place gel on the UV box</li>
-	<li> Carefully extract the fragment suspended in the gel/li>
+	<li> Carefully extract the fragment suspended in the gel </li>
 	<li> Mass gel fragments </li>
 	<li> Place fragment into a 1.5 mL tube and add 4 &micro;L of H2O </li>