Technical Data
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5'-T^G A T C A-3'
3'-A C T A G^T-5'

Concentration: 10u/ul

Source: Bacillus caldolyticus

Buffer: 10mM Tris-HCl (pH 7.5), 10mM MgCl2, 50mM NaCl and 0.1mg/ml BSA. Incubate at 55C. Incubation at 37C results in 50% activity.

Double Digests Y+/Tango Buffer is provided to simplify buffer selection for double digests. 98% of Fermentas Restriction Endonucleases (REs) work well in a 1X or 2X concentration of the Y+/Tango Buffer. Please refer to the Enzyme Activity Chart to choose the best buffer for the two REs in your digest.

Diluent Buffer: 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.

Storage Buffer: 10mM Tris-HCl (pH 7.4 at 25C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion Assay: No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with BclI (see Star Activity).

Ligation/Recutting Assay: After 50-fold overdigestion (3u/ug DNA x 17 hours) with BclI, more than 90% of the DNA fragments can be ligated at a 5'-termini concentration of 0.1uM. More than 90% of these can be recut.

Labeled Oligonucleotide (LO) Assay: No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10units of restriction endonuclease for 4 hours.

Blue/White Cloning Assay: The mix of pUC57/HindIII, pUC57/PstI and pUC57/Eco32I digests was incubated with 10units of enzyme for 16 hours. After religation and transformation 0.3% of white colonies were detected.

Star Activity: A large excess of enzyme (7.5u/ug DNA x 16 hours) may result in star activity. Star activity appears at 10-fold overdigestion (10 units x 1 hour).

Methylation Effects: BclI cuts neither TGm6ATCA nor TGAThm5CA, but cuts TGATm5CA.

Blocked by Dam methylation.

Stability during Prolonged Incubation: A minimum of 0.1unit of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 55C.

Thermal Inactivation: Only small amounts of enzyme (up to 10units) can be inactivated at 80C for 20min.

Compatible Ends: BamHI, BglII, Bsp143I, MboI, PsuI

Number of Recognition Sites in DNA:
Lambda: 8
PhiX174: 0
M13mp18/19: 0
pBR322: 0
pUC18/19: 0
pUC57: 0
pTZ19R/U: 0
pBluescriptIIKS(-/+): 0
pBluescriptIISK(-/+): 0
pACYC177: 0
pACYC184: 1

Unit Definition: One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dam-in 1 hour at 55C in 50ul of assay buffer.
Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.