Product info: PspC I


Name
PspC I
Cat. #E475TE476T
Number of reactions100500
Volume, μl100500

Recognition site
CACGTG
GTGCAC
SourcePseudomonas species C
Assayed onLambda DNA, plasmid DNA
DescriptionTurbo PspC I is used for short time (10 min) DNA digestion in SE-Buffer B+.
Applications:
- Fast DNA analysis
- Fast preparation of vectors for cloning
- Double digestion
Reaction bufferSE-buffer B+ (10 mM Tris-HCl (pH 7.6 at 25°C); 10 mM MgCl2; 1 mM DTT; 100 μg/ml BSA .)
Optimal temperature
37oC
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.
Storage temperature: -20°C.
Storage at -70°C is recommended for periods longer than 30 days.
LigationAfter digestion with 1 μl of Turbo PspC 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 B+
Methylation sensitivitynot tested
Inactivation 20 minutes under
65oC
NotesEnzyme Properties:
1 μl of Turbo PspC I cuts 1 μg of DNA in 1x SE-Buffer B+ 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 B+ - 2 μl
DNA - 0.2-1 μg
Nuclease-free water - to 20 μl
+ 1 μl of Turbo PspC I
Mix by pipette tip carefully.
Incubate at 37°C for 10 min.
References:Tronin, A., Kileva, E.V., Mezentseva, N., Dedkov, V.S., Degtyarev, S.K. Unpublished observations. (2003)