Recognition & cleavage sequence

Source: Brevibacterium albidum

Reaction conditions

1X EzBuffer Bal I, 37℃

1X EzBuffer Bal I

50 mM Tris-HCI (pH 8.2 @ 25℃), 5 mM MgCl₂

Unit definition

One unit is defined as the amount of enzyme required for complete digestion of 1-μg bacteriophage λ at 37℃ for 1 hr in 50-μl reaction mixtures.

Storage

10 mM Tris-HCl (pH 7.4 @ 25℃), 50 mM NaCl, 0.1 mM EDTA, 1 mM dithiothreitol, 500 μg/ml BSA, 50% glycerol.

Dilution buffer: EzDiluent B

10 mM Tris-HCl (pH 7.4 @ 25℃), 300 mM NaCl, 0.1 mM EDTA, 1 mM dithiothreitol, 500 μg/ml BSA, 50% glycerol.

Heat Inactivation

Bal I can be inactivated at 65℃ for 20 min.

Methylation sensitivity

dam methylation: Not sensitive

dcm methylation: Conditionally sensitive

CpG methylation: Not sensitive

Relative activity in EzBuffers

EzBuffer I: 0%

EzBuffer II: 75%

EzBuffer III: 25%

EzBuffer IV: 75%

FastCut Buffer: Not recommended

Note

Activity is inhibited by dcm methylation partially overlapping its recognition sequence.

●  Enzynomics_R056_Bal I_manual

● Material Safety Data Sheet

- Enzynomics_MSDS_R056_Bal I

● Quality control

Overdigestion assay

DNA digested for 16 hr in 50-μl reaction with 100 U of enzyme resulted in same the DNA band patterns as those obtained with 1 U of enzyme for 1 hr.

Ligation and recutting

More than 95% of DNA fragments (1 μg) digested with 20-fold excess enzyme can be ligated by T4 DNA ligase (400 U) at 16℃ for 16 hr. Of the ligation products, > 95% can be re-cut. 

Extreme pure (EP)

No detectable degradation of ³²P-end labeled single-stranded and double-stranded (5’-, 3’-overhang and blunt end) oligonucleotides occurred during incubation with 100 U of enzyme for 4 hr.