Team:Newcastle/Gel extraction

From 2010.igem.org

QIAquick gel extraction microcentrifuge protocol

1. Excise the DNA fragment from the agarose gel with a clean, sharp scalpel
2. Weight the gel slice and add 3 volumes of buffer QG to 1 volume of gel (100 mg ~ 100 µl)
3. Incubate at 50°C and invert the tube gently at regular intrerval until the gel has completely dissolved
4. After the gel has dissovled completely, check that the color of the mixture is yellow
5. Add 1 gel volume of isopropanol to the sample and mix
6. Place a QIAquick spin column in a 2 ml collection tube
7. To bind DNA, apple the sample to the QIAquick column and centrifuge for 1 min
8. Discard the flow through and place the QIAquick column back into the same tube (Maximum volumn is 750ul)
9. Add 0.5 ml of Buffer QG to QIAquick column and centrifuge for 1 min and discard the flow through and place the QIAquick column back into the same tube
10. To wash, add 0.75 ml of Buffer PE to QIAquick column and centrifuge for 1 min and place the QIAquick column back into the same tube
11. Centrifuge the column for a further 1 min
12. Transfer the column into a clean 1.5 ml micriocentrifuge tube
13. To elute DNA, add 30 µl of Buffer EB to the center of the QIAquick membrane and allow to stand for 1 min
14. Centrifuge the column for 1 min and transfer the eluate to a clean 1.5 ml micriocentrifuge tube