I used the MNase from NEB, which came with its own buffer. 1x NEB reaction buffer is 50 mM Tris-HCl, 5 mM CaCl2, which should result in pH 7.9 at 25degC. Additionally, the reaction should be supplemented with 1X NEB BSA (100 ug/ml).
For me, this worked very well. 5000 units (2.5 ul) digest 50 ug of chromatin in a range from mainly mono-penta nucleosomes in one minute.
I used the MNase from NEB, which came with its own buffer. 1x NEB reaction buffer is 50 mM Tris-HCl, 5 mM CaCl2, which should result in pH 7.9 at 25degC. Additionally, the reaction should be supplemented with 1X NEB BSA (100 ug/ml).
For me, this worked very well. 5000 units (2.5 ul) digest 50 ug of chromatin in a range from mainly mono-penta nucleosomes in one minute.
Hi! I'm trying to process genomic DNA of Phytophthora infestans, and it is not working!!
I've used 10 ug of DNA, 1000 units, 1x buffer and 1x BSA of New England Biosciences micrococcal nuclease! With different processing times of 1, 3, 5, 10, 15 min at 37ºC. Both with and without EDTA (10mM) to stop the reaction and nothing works!
I've then loaded the samples to a 1% agarose gel but I don't see anything other than the ladder.
HELP!
- Badges
- Science topic
- Similar topics
- Biological Science
- Cell Biology