Product info: Psp124B I


Name
Psp124B I
Cat. #E107TE108T
Number of reactions50250
Volume, μl50250

Recognition site
GAGCTC
CTCGAG
SourcePseudomonas species 124B
Assayed onLambda DNA (HindIII-digest), plasmid DNA
DescriptionTurbo Psp124B I is used for short time (10 min) DNA digestion in SE-Buffer G.
Applications:
- Fast DNA analysis
- Fast preparation of vectors for cloning
- Double digestion
Reaction bufferSE-buffer G (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 50 mM NaCl; 1 mM DTT.)
Optimal temperature
37oC
Storage conditions10 mM Tris-HCl (pH 7.5); 250 mM NaCl; 0,1 mM EDTA; 7 mM 2-mercaptoethanol; 100 μg/ml BSA; 50% glycerol; Store at -20°C.
LigationAfter digestion with 1 μl of Turbo Psp124B 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 G.
Methylation sensitivitynot tested
Inactivation 20 minutes under
80oC
NotesEnzyme Properties:
1 μl of Turbo Psp124B I cuts 1 μg of DNA in 1x SE-Buffer G 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 G - 2 μl
DNA - 0.2-1 μg
Nuclease-free water - to 20 μl
+ 1 μl of Turbo Psp124B I
Mix by pipette tip carefully.
Incubate at 37°C for 10 min.
References:Degtyarev, S.Kh., Dedkov, V.S., Rechkunova, N.I., Morgan, R. Unpublished observations (1994).