| 5'-G^G A T C C-3' |
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| 3'-C C T A G^G-5' |
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| Catalog # | B0070 |
| Source | Bacillus amyloliquefaciens H |
| Supplied with | B0070A: Reaction Buffer (10X) for BamHI+, 10X |
| Supplied as a liquid in10mM Tris-HCl, pH 8.0, 5mM MgCl2, 100mM KCl, 0.02% Triton X-100, 0.1mg/ml BSA. Supplied with 2x1ml of Reaction Buffer (10X). |
| Concentration | 10u/ul |
| Unit Definition | One unit is defined as the amount of enzyme required to digest 1ug of lambda DNA-Bsp120I fragments in 1 hour at 37°C in 50ul of assay buffer. |
| Diluent Buffer | 10mM Tris-HCl, pH 7.4, 100mM KCl, 1mM EDTA, 1mM DTT, 0.2mg/ml BSA, 50% glycerol. |
| Storage Buffer | 10mM Tris-HCl, pH 7.4 at 25ºC, 200mM NaCl, 1mM DTT, 1mM EDTA, 0.2mg/ml BSA, 50% glycerol. |
| Incubation Temp | 37°C |
| Methylation Effects | BamHI does not cut GGATm4CC, GGATm5CC, GGAThm5Chm5C, GGAhm5UCC, but cuts GGATCm5C, GGm6ATCC, GGm6ATCm5C and GGATCm4C. |
| Number of Recognition Sites in DNA Molecules | Lambda: 5 |
| PhiX174: 1 |
| M13mp18/19: 1 |
| pBR322: 1 |
| pUC18/19: 1 |
| pUC57: 1 |
| pTZ19R/U: 1 |
| Stability during Prolonged Incubation | A minimum of 0.5 units of enzyme are required for complete digestion of 1ug of lambda DNA in 16 hours at 37°C. |
| Thermal Inactivation | Only small amounts of enzyme (up to 10 units) can be inactivated at 80°C for 20min. |
| Compatible Ends | BclI, BglII, Bsp143I, MboI, PsuI |
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| Overdigestion Assay | |
| No detectable change in the specific fragmentation pattern is observed after 80-fold overdigestion (5u/ug lambda DNA x 16 hours) with BamHI (see Star Activity). |
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| Ligation/Recutting Assay | |
| After 50-fold overdigestion (3u/ug DNA x 17 hours) with BamHI, more than 95% of the DNA fragments can be ligated at a 5'-termini concentration of 0.02uM. More than 95% of these can be recut. |
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| Labeled Oligonucleotide (LO) Assay | |
| No detectable degradation of a single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10 units of restriction endonuclease for 4 hours. |
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| Blue/White Cloning Assay | PUC57 was digested at a unique site with 10 units of enzyme for 16 hours. After religation and transformation 0.2% of white colonies were detected. |
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| Digestion of Agarose-embedded DNA | |
| A minimum of 5 units of enzume is required for complete digestion of 1ug of agarose- embedded lambda DNA in 16 hours. |
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| Star Activity | |
| Appears at > 5-fold overdigestion at (5u x 1h). |
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