Product info: Sfr274 I


Name
Sfr274 IBlocked by 
 CTCG(6mA)G methylation  Not blocked by 
CT(5mC)GAG methylation  
Cat. #E125TE126T
Number of reactions100500
Volume, μl100500

Recognition site
CTCGAG
GAGCTC
SourceStreptomyces fradiae 274
Assayed onLambda DNA (Hind III-digest), plasmid DNA
DescriptionTurbo Sfr274 I is used for short time (10 min) DNA digestion in universal (ROSE) SE-Buffer.
Reaction Original SibEnzyme (ROSE) Buffer is a specially designed universal reaction buffer for the most Restriction Endonucleases. ROSE Buffer is perfect for DNA cleavage with SE Turbo Restriction Endonucleases and for double digestion.
Applications:
- Fast DNA analysis
- Fast preparation of vectors for cloning
- Double digestion
Reaction bufferSE-buffer ROSE
Optimal temperature
50oC
Storage conditions10 mM Tris-HCl (pH 7.5); 50 mM KCl; 0.1 mM EDTA; 7 mM 2-mercaptoethanol; 200 μg/ml BSA; 50% glycerol.. Store at -20°C.
LigationAfter digestion with 1 μl of Turbo Sfr274 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
Methylation sensitivityBlocked by CTCGmAG methylation.
Not blocked by CTmCGAG methylation.
Inactivation 20 minutes under
65oC
NotesEnzyme Properties:
1 μl of Turbo Sfr274 I cuts 1 μg of DNA in 1x SE-Buffer ROSE in 10 min (assayed on Lambda/HindIII 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 Sfr274 I
Mix by pipette tip carefully.
Incubate at 50°C for 10 min.
Incubation at 37°C rusults in 75-100% activity.
References:Puchkova, L.I., Krivopalova, G.N., Andreeva, I.S., Selina, A.V., Serov, G.D., Rechkunova, N.I., Degtyarev, S.Kh. Izv. Sib. Otd. Akad. Nauk SSSR 1: 32-34 (1990).