New England Biolabs
AflII
R0520S
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I have constructed a plasmid called pUC19_atpA of size 5113 bp. These constructed were transformed into E. coli DH5 alpha cells. Plasmid was prepped from E. coli DH5 alpha cells to quickly confirm presence of this plasmid by diagnostic digestion with AflII restriction enzyme from NEB. Restriction with AflII was expected two give DNA fragments of 4192 and 921 bp size. Indeed, restriction of the plasmid prepped gave correct restriction pattern as seen in 1 % agarose gel.
Diagnostic restriction mapping of the plasmid constructs
1 microgram of the plasmid DNA
Restriction digestion reactions were set up in a 50 microliter reaction volume with 1 microgram plasmid DNA prepped (plasmid mini preps), NEB's Cutsmart buffer to a final concentration of 1X, and 1 microliter of AflII restriction enzyme. Digestion mix was incubated at 37 degree C for 1 hour and then analysed by 1% agarose gel run.
Plasmid DNA minipreps should be of high quality without any high salt contamination. Otherwise inhibition of AflII is possible.
Restriction digestion of the plasmid construct, pUC19_atpA with AflII enzyme gave expected restriction pattern of 4192 and 921 bp DNA fragments as visualised by 1% agarose gel run.
None
Easy and simple protocol.
Diagnostic restriction mapping of the my plasmid construct with AflII restriction enzyme from NEB went well and satisfied using this product for my molecular cloning experiments.