Previously added items:
5'...G↓G T N A C C...3'3'...C C A N T G↑G...5'
Lambda DNA digested with Eco91I, 0.7% agarose, 13 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.
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.
G↓GTSACC
G↓GTWACC
1X FastDigest buffer or 1X FastDigest Green buffer at 37°C. 1 µL of FastDigest Eco91I is formulated to digest up to:
(CCWGG)
5'...Cm5CW GGTNACm5CW GG...3'
3'...G GWm5CCANTG GWm5CC...5'
5'...m5C GGTNACm5C G...3'
3'... Gm5CCANTG Gm5C...5'
G^GTNACC
GGTNACGTNACC
G^GTAACC
GGTAACGTAACC
G^GTCACC
GGTCACGTCACC
G^GTGACC
GGTGACGTGACC
G^GTTACC
GGTTACGTTACC
GACN^NNGTC
GACNNNNGTC
GACN^ANGTC
GACNAANGTC
GACN^CNGTC
GACNCCNGTC
GACN^GNGTC
GACNGGNGTC
GACN^TNGTC
GACNTTNGTC