New England Biolabs
AhdI
R0584S
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I have constructed a plasmid of size around 7.2 Kb and transformed into appropriate E. coli cells. Then plasmid prepped form these cells were used for diagnostic restriction mapping using NEB's AhdI enzyme. My plasmid construct has restriction site for AhdI and will give two bands of 6 and 1.2 kb respectively. Restriction digestion and agarose gel analysis of the plasmid construct showed correct restriction mapping.
Restriction mapping and cloning
1 microgram of plasmid DNA
The protocol is very simple. Set up reactions in a 50 microliter volumes with 1 microgram of plasmid DNA, appropriate buffer and 1 microliter of AhdI enzyme. Incubate reaction at 37 degree C for 1 hour and then perform 1% agarose gel run.
Use appropriate buffer system with chosen enzyme.
Restriction digestion of my plasmid construct gave two bands of 6 an 1.2 kb respectively. This is the expected Restriction map as shown in agarose gel.
N/A
None
Simple and easy protocol
Restriction mapping of my plasmid construct went well. I recommend using this enzyme for restriction mapping of the plasmids if the plasmid have AhdI restriction site.