Previously added items:
5'...G A A G A (N)8↓...3'3'...C T T C T (N)7↑...5'
Lambda DNA (dam-) digested with MboII, 1.4% agarose, 130 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 lambda DNA (dam) (#SD0021). MboII is blocked by overlappingdam methylation. To avoid dam methylation, use a dam, dcm- strain. Greater than 15-fold overdigestion with MboII may result in star activity. MboII produces DNA fragments that have a single-base 3 extension, which are more difficult to ligate than blunt-ended fragments.
MboII may remain associated with the cleaved DNA. This may cause DNA band shifting during electrophoresis. To avoid atypical DNA band patterns, use the 6X DNA Loading Dye & SDS Solution for sample preparation or heat the digested DNA in the presence of SDS prior to electrophoresis.
For methylation sensitivity refer to product specifications.
GATC
GAAGm6A TC
CTTC Tm6AG
m5C GAAGA
Gm5CTTCT
TGA(N)8TGCT
TGm6AAGA(N)5 TGCT
AC TTCT(N)5m6ACGA