"

From 2010.igem.org

Gram-positive Bacteria Chromosomal DNA Extraction Protocol

Puregene DNA isolation

1. Grow overnight 7.5ml cultures in THYB containing 30μg/ml hyaluronidase.

Cell Lysis

1. Cells go through 3600 x g centrifugation for 15 minutes to pellet.
2. The supernatant is poured away.
3. 0.5ml of cell suspension solution is added to the eppendorf tube. The solution in the tube is pipetted up and down to resuspend cells. The solution is then transferred to another 1.5ml tube.
4. 50μl of lysozyme and 7.5μl of lytic enzyme solution is added into the tube. The tube is then inverted 25 times to mix the solution.
5. We then incubate it at 37°C