Team:Sevilla/Week5

Purification of DNA from agarose gel

GenElute Gel Extraction Kit from SIGMA

1.Excise band fron the agarose gel and weigh the gel

2.For every 100 mg of agarose gel, add 300 μl of Gel Solubilization Solution. Incubate the gel mixture at 55ºC for 10 minutes, or until the gel slice is completely dissolved. Vortex every 2-3 minutes.

3.Prepare binding column. Place the GenElute Binding Column G into one of 2 ml collection tubes. Add 500 μl of the Column Preparation Solution to each binding column. Centrifuge for 1 minute.

4.Add 1 gel volume of 100% isopropanol and mix until homogenous.

5.Load the solubilized gel solution mixture into the binding column. Centrifuge 1 min. Discard the flow-through

6.Add 700 μl of Wash Solution to the binding column. Centrifuge for 1 minute. Discard the flow-through and repeat this step to remove excess ethanol.

7.Transfer the binding column to a fresh collection tube. Add 30 μl of Elution Solution to the center of the membrane and incubate for 1 minute. Centrifuge for 1 min.