Product info: Mfe I


Name
Mfe I
Cat. #E295TE296T
Number of reactions50250
Volume, μl50250

Recognition site
CAATTG
GTTAAC
SourceAn E.coli strain, that carries the cloned gene Mfe I from Mycoplasma fermentans.
Assayed onLambda DNA, plasmid DNA
DescriptionTurbo Mfe I is used for short time (10 min) DNA digestion in universal (ROSE+) SE-Buffer.
Reaction Original SibEnzyme (ROSE+) Buffer is a modified universal ROSE (Reaction Original SibEnzyme) Buffer, specially designed for Restriction Endonucleases that require BSA to obtain 100% activity. The concentration of BSA in 1X ROSE+ Buffer is 100 μg/ml. ROSE+ Buffer is perfect for DNA cleavage with SE Turbo Restriction Endonucleases and for double digestion. Do not use this Buffer for long incubation (16 hours) with the enzymes having star activity.
Applications:
- Fast DNA analysis
- Fast preparation of vectors for cloning
- Double digestion
Reaction bufferSE-buffer ROSE+
Optimal temperature
37oC
Storage conditions10 mM Tris-HCl (pH 7.6); 50 mM NaCl; 0,1 mM EDTA; 1 mM DTT; 200 μg/ml BSA; 50% glycerol. Store at -20°C.
LigationAfter digestion with 1 μl of Turbo Mfe I, approximately 90% of the DNA fragments can be ligated with high-activity T4 DNA Ligase and recut.
Non-specific hydrolisisNo detectable degradation of a single-stranded and double-stranded oligonucleotide was observed after incubation with 1 μl of restriction endonuclease for 3 hours.
Reagents Supplied with Enzyme 10 X SE-buffer ROSE+
Inactivation 20 minutes
No
Notes1 μl of Turbo Mfe I cuts 1 μg of DNA in 1x SE-Buffer ROSE+ in 10 min (assayed on Lambda and plasmid DNA). A short time of DNA digestion requires high quality purification of DNA sample (PCR fragments should be purified after amplification).
Please note that supercoiled plasmid DNA and PCR fragments may have varying rates of cleavage and sometimes need more time to be completely digested.
Standard protocol of Turbo reaction:
20 μl of the reaction volume:
10x SE-Buffer ROSE+ - 2 μl
DNA - 0.2-1 μg
Nuclease-free water - to 20 μl
+ 1 μl of Turbo Mfe I
Mix by pipette tip carefully.
Incubate at 37°C for 10 min.