NotI NEB#R0189

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Methylated DNAs were purified and concentrated by Phenol/Chloroform extraction and ethanol precipitation.	The extent of protection was determined by cleavage using HaeIII and NotI (NEB) restriction enzymes according to the supplier's instructions.

Upstream tips
Methylated DNAs were purified and concentrated by Phenol/Chloroform extraction and ethanol precipitation.
Protocol tips
The extent of protection was determined by cleavage using HaeIII and NotI (NEB) restriction enzymes according to the supplier's instructions.

Publication protocol

For in vitro methylation, DNA isolated from E. coli DH5α (dam+dcm+) was treated with either M.CbeI or M.HaeIII methyltransferase (NEB). 5 microgram of purified M.CbeI was incubated with 50 mM Tris-HCl, 50 mM NaCl, 80 μM S-adenosylmethionine (SAM), 10 mM Dithiothreitol (DTT) at pH 8.5 and 20 µg of DNA substrate in 400 µl reaction, and incubate for 2 hours at 78°C. The M.HaeIII methylation reaction was performed according supplier's instructions. To allow complete methylation, an additional 10 units of M.HaeIII and 80 µM SAM was added to the reaction every 4 h of incubation at 37°C for a total of 12 h. Methylated DNAs were purified and concentrated by Phenol/Chloroform extraction and ethanol precipitation. The extent of protection was determined by cleavage using HaeIII and NotI (NEB) restriction enzymes according to the supplier's instructions.

Full paper   Login or join for free to view the full paper.

Reviews

Discussion

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing Restriction Enzymes NotI using NotI NEB#R0189 from New England BioLabs.

Manufacturer protocol

Download the product protocol from New England BioLabs for NotI NEB#R0189 below.

Videos

Check out videos that might be relevant for performing Restriction Enzymes NotI using NotI NEB#R0189 from New England BioLabs. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.