The 1-base overhangs produced by PflFI may be hard to ligate.Sticky ends from different PflFI sites may not be compatible.
Tth111I (5569) 1 site
GACNNNGTCCTGNNNCAG
The 1-base overhangs produced by Tth111I may be hard to ligate.Sticky ends from different Tth111I sites may not be compatible.
BseRI (5221) 1 site
GAGGAG(N)8NNCTCCTC(N)8
Sticky ends from different BseRI sites may not be compatible.BseRI quickly loses activity at 37°C.Prolonged incubation with BseRI may lead to degradation of the DNA.
BmtI (5097) 1 site
GCTAGCCGATCG
NheI (5093) 1 site
GCTAGCCGATCG
NdeI (4824) 1 site
CATATGGTATAC
Prolonged incubation with NdeI may lead to removal of additional nucleotides.