HaeIII NEB#R0108

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	The extent of protection was determined by cleavage using HaeIII and NotI (NEB) restriction enzymes according to the supplier's instructions.

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.

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