Previously added items:
5'...G G C C N N N N↓N G G C C...3'3'...C C G G N↑N N N N C C G G...5'
Ad2 DNA digested with SfiI, 0.7% agarose, 3 cleavage sites
Thermo Scientific conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.
Assayed using pUC19 DNA with insert containing two SfiI sites from Adenovirus-2 DNA. Incubation at 37°C results in 10% activity. SfiI cleavage is impaired by overlapping dcm methylation. To avoid dcm methylation, use a dam-, dcm- strain, such as GM2163.
For cleavage with SfiI, at least two copies of its recognition sequence are required. The two sites can be on either the same or different DNA molecules. (L. M. Wertzell et al., J. Mol. Biol., 248, 581-595, 1995.) Therefore, also an oligonucleotide harboring a SfiI recognition site can be supplemented.
For methylation sensitivity refer to product specifications.
Cm5CW GGCC(N)5GGCm5CW GG
G GWm5CCGG(N)5CCG GWm5CC
m5C GGCC(N)5GGCm5C G
GGCm5C G(N)3m5C GGCC
CCG Gm5C(N)3 Gm5CCGG