Fragment Purification

Fragment Purification from an Agarose Gel

1. Add 5X loading buffer to the restriction digest (eg. 40 ul digest + 10 ul 5X) and run on 1% Agarose gel
2. With a plastic ruler, and wearing safety glasses, slice the band out of the gel on the UV box
3. Weigh the gel slice (expect 100-250 mg)
4. Use the Nucleospin Extract IIkit from Machery-Nagel
5. Add 2 volumes of buffer NT
6. Incubate 5 minutes at 50 C, vortexing 2 times along the way and at the end
7. Transfer into a Nucleospin Extract II column on a collection tube and spin 1 minute at full speed
8. Dump collection tube and add 700 ul buffer NT3 to column
9. Spin 30 sec at full speed