Previously added items:
5'...C T G A A G (N)16↓...3'3'...G A C T T C (N)14↑...5'
Lambda DNA digested with AcuI, 1% agarose, 40 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.
FastDigest AcuI (Eco57I) requires only Mg2+ for its activity, but is stimulated by SAM. 0.01 mM SAM gives more than a 100-fold increase in FastDigest AcuI (Eco57I) activity. Still, a complete cleavage of some substrates with FastDigest AcuI (Eco57I) is difficult to achieve. For cleavage with FastDigest AcuI (Eco57I), at least two copies of its recognition sequence are required.
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 + 0.01 mM SAM at 37°C. 1 µl of FastDigest AcuI (Eco57I) is formulated to digest up to: