Previously added items:
5'...A C C T G C (N)4↓...3'3'...T G G A C G (N)8↑...5'
Lambda DNA digested with BspMI (BveI), 0.7% agarose, 41 cleavage sites
Thermo Scientific FastDigest enzymes are an advanced line of fast restriction enzymes that are all 100% active in the universal FastDigest and FastDigest Green reaction buffers.
The universal buffer allows rapid single-, double- or multiple DNA digestion within 5-15 minutes eliminating any need for buffer change or subsequent DNA clean-up steps. DNA modifying enzymes, such as Klenow Fragment, T4 DNA Ligase, alkaline phosphatases and T4 DNA Polymerase all have 100% activity in FastDigest Buffer. Therefore, enzymes for downstream applications can be directly added to the FastDigest reaction mix.
For additional convenience FastDigest Green Buffer includes a density reagent and two tracking dyes for direct loading of digestion reaction products on gels.
Short incubation times and optimal composition of the universal FastDigest Buffer eliminate star activity effects.
At least two copies of FastDigest BspMI (BveI) recognition site are required for efficient cleavage. Inclusion of 0.5 µM oligonucleotide with the FastDigest BspMI (BveI) recognition sequence in the reaction mixture significantly improves cleavage of plasmid DNAs, especially of those with a single FastDigest BspMI (BveI) site. Still, a complete cleavage of some substrates by FastDigest BspMI (BveI) is difficult to achieve.
The FastDigest Green Buffer offers the same high performance in DNA digestion and downstream applications as the colorless FastDigest Buffer. For applications that require product analysis by fluorescence excitation (e.g. concentration measurements in UV light) the colorless FastDigest Buffer is recommended.
For methylation sensitivity refer to product specifications.
1X FastDigest buffer or 1X FastDigest Green buffer + 0.5 µM oligonucleotide at 37°C. 1 µL of FastDigest BspMI (BveI) is formulated to digest up to: