After lysis of bacterial cell whenever I went for centrifugation every time my supernatant(have protein of interest) got mixed with pellet even centrifugation at very high (g) and low temperature also.
need your kind suggestion
Hello Asmat Ullah
You can use Micrococcal Nuclease (MNase) which is suitable for removing nucleic acids from cell lysates. Please go through the link below for more details on Micrococcal Nuclease.
https://www.thermofisher.com/order/catalog/product/88216
Also, you can refer to the research article attached. It may be helpful.
Article Removal of nucleic acid contaminants using nuclease enzymes ...
Hope this helps.
Best.
You can use DNAse or benzonase to reduce the prep viscosity and cut large DNA molecules after that you can add 1M NaCl to remove nucleic acid.
Alternative, you can change the PEI percentage maybe 0.05% final (first centrifuge as normal then add PEI to the supernatant at 0.05 % final stir for 20 min And then centrifuge it at 31,000 xg, 4C for 30-45 min )
- Badges
- Science method