Previously added items:
5'...C A C C T G C (N)4↓...3'3'...G T G G A C G (N)8↑...5'
Lambda DNA digested with AarI, 1% agarose, 12 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.
For methylation sensitivity refer to product specifications.
For cleavage with AarI at least two copies of its recognition sequence are required. Inclusion of 0.5 µM oligonucleotide with the AarI recognition sequence in the reaction mixture significantly improves cleavage of DNAs, especially of those with a single AarI site.
Still, a complete cleavage of some substrates with AarI is difficult to achieve. Greater than 10-fold overdigestion with AarI may result in star activity. AarI 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.