Product info: HspA I


Name
HspA IBlocked by 
5`-G(5mC)GC-3`/3`-CG(5mC)G-5` methylation  Not blocked by  
5`-GCG(5mC)-3`/3`-CGCG-5` methylation    
Cat. #E069TE070T
Number of reactions50250
Volume, μl50250

Recognition site
GCGC
CGCG
SourceAn E.coli strain that carries the cloned HspA I gene from Haemophilus species A1
Assayed onLambda DNA, plasmid DNA
DescriptionTurbo HspA 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
37oC
Storage conditions10 mM Tris-HCl (pH 7.5); 50 mM NaCl; 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 HspA 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 CG methylation 5'-G(5mC)GC-3'/3'-CG(5mC)G-5'.
Not blocked by methylation 5'-GCG(5mC)-3'/3'-CGCG-5'.
Cut hemimethylated site: 5'- G(5mC)GC-3'/3'-CGCG-5`
Inactivation 20 minutes under
80oC
NotesEnzyme Properties:
1 μl of Turbo HspA 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 HspA I
Mix by pipette tip carefully.
Incubate at 37°C for 10 min.
References:Rechkunova, N.I., Prikhod'ko, E.A., Shevchenko, A.V., Degtyarev, S.K. Unpublished observations (1994).
 Chernukhin V.A, Abdurashitov M.A., Tomilov V.N., Gonchar D.A., Degtyarev S.Kh. Comparative analysis of mouse chromosomal DNA digestion with restriction endonucleases in vitro and in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.3, No 4, pp 19-27, 2007
 V.A. Chernukhin, M.A. Abdurashitov, V.N. Tomilov, D.A. Gonchar, S.Kh. Degtyarev Comparative restriction enzymes analysis of rat chromosomal DNA in vitro and in silico // "Ovchinnikov bulletin of biotechnology and physical and chemical biology" V.2, No 3, pp 39-46, 2006