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'
pUC19 DNA with inserted SfiI recognition sites, 0.7% agarose, 2 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.
For cleavage with FastDigest SfiI, 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 at 50°C. 1 µL of FastDigest SfiI is formulated to digest up to:
5'...Cm5CW GGCC(N)5GGCm5CW GG...3'
3'...G GWm5CCGG(N)5CCG GWm5CC...5'
5'...m5C GGCC(N)5GGCm5C G...3'
3'... Gm5CCGG(N)5CCG Gm5C...5'
5'...GGCm5C G(N)3m5C GGCC...3'
3'...CCG Gm5C(N)3 Gm5CCGG...5'