Login

Forgot your password?
New User?
Remember me
banner banner

You are here:Home » Molecular Biology » MB-Enzymes, Restriction » BclI

BclI

Pricing

  For pricing information, USA customers sign in.
  Outside USA? Please contact your distributor for pricing.

Specifications

5'-T^G A T C A-3'
3'-A C T A G^T-5'
Catalog #B0807-01
Concentration 10u/ul
Source Bacillus caldolyticus
Buffer 10mM Tris-HCl (pH 7.5), 10mM MgCl2, 50mM NaCl and 0.1mg/ml BSA. Incubate at 55°C. Incubation at 37°C 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 25°C), 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA and 50% glycerol.
Storage Buffer 10mM Tris-HCl (pH 7.4 at 25°C), 100mM KCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA and 50% glycerol.
Overdigestion AssayNo 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 AssayAfter 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) AssayNo 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 AssayThe 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 ActivityA 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 EffectsBclI cuts neither TGm6ATCA nor TGAThm5CA, but cuts TGATm5CA.
Blocked by Dam methylation.
Stability during Prolonged IncubationA minimum of 0.1unit of enzyme is required for complete digestion of 1ug of lambda DNA in 16 hours at 55°C.
Thermal InactivationOnly small amounts of enzyme (up to 10units) can be inactivated at 80°C for 20min.
Compatible EndsBamHI, BglII, Bsp143I, MboI, PsuI
Number of Recognition Sites in DNA
Lambda8
PhiX1740
M13mp18/190
pBR3220
pUC18/190
pUC570
pTZ19R/U0
pBluescriptIIKS(-/+)0
pBluescriptIISK(-/+)0
pACYC1770
pACYC1841
Unit DefinitionOne unit is defined as the amount of enzyme required to digest 1ug of lambda DNA dam-in 1 hour at 55°C in 50ul of assay buffer.


External Links