What's up with Afl II and Bfr I?

John Dixon jpcd0 at mole.bio.cam.ac.uk
Mon Nov 18 08:49:37 EST 1996

Hi there, has anyone had experience with Afl II (NEB) or Bfr I (BM)?

They are isoschizomers cutting the site C/TTAAG but NEB warn that less
than 50% fragments religate under standard conditions and recommend using
High Conc ligase. Their enzyme passes the blue white assay, so it's not
nibbling ends. Perhaps it is something intrinsic to a TTAA 5' extension;
no other enzyme cuts to leave such an end, although EcoRI ends (AATT 5')'
ext are clearly not problematic.

Boerhinger say that they are not aware of any problem with their
equivalent, BfrI. Has anyone used this for cloning?

Any clues on Afl II end problems?



John Dixon                                        Lab 44 (1223) 334131
Wellcome/CRC Institute                            Fax 44 (1223) 334134
Department of Genetics
Cambridge University    
United Kingdom                           e-m: jpcd0 at mole.bio.cam.ac.uk

More information about the Methods mailing list