Bln I (Avr II)
ROCHE/BLNI-RO - from Brevibacterium linens
Product Type: Chemical
Bln I (Avr II)
| application(s) | life science and biopharma sample preparation |
| biological source | bacterial (Brevibacterium linens) |
| color | colorless |
| foreign activity | Endonucleases, none detected (up to 20 U with MWM II-DNA) |
| Endonucleases, none detected (up to 20U with pBR 322-DNA) | |
| form | solution |
| manufacturer/tradename | Roche |
| packaging | pkg of 1,000 U (11558170001 [10 U/μl]) |
| pkg of 200 U (11558161001 [10 U/μl]) | |
| parameter | 37 °C optimum reaction temp. |
| pH | 8.1 (39 °F) |
| Quality Level | 100  |
| shipped in | dry ice |
| solubility | water: miscible |
| specific activity | 10000 U/mL |
| storage temp. | −20°C |
| suitability | suitable for molecular biology |
| Analysis Note: | PFGE tested Bln I has been tested in Pulsed-Field Gel Electrophoresis (on bacterial chromosomes). For cleavage of genomic DNA (E.coli C 600) embedded in agarose for PFGE analysis, we recommend using 10 U of enzyme/μg DNA and 4 hour incubation. |
| Analysis Note: | SuRE/Cut Buffer System The buffer in bold is recommended for optimal activity • A: 25-50% • B: 50-75% • H: 100% • L: 0-10% • M: 25-50% |
| Analysis Note: | Activity in PCR buffer: Not tested |
| DNA Profile: | Number of cleavage sites on different DNAs • λ: 2 • φX174: 0 • Ad2: 2 • M13mp7: 0 • M13mp18:0 • pBR322: 0 • pBR328: 0 • pUC18: 0 • SV40: 2 |
| General description: | Bln I recognizes the sequence C↓CTAGG and generates fragments with 5′-cohesive termini. Compatible ends Bln I ends are compatible with ends generated by Nhe I, Spe I and Xba I. Isoschizomers Bln I is an isoschizomer of Avr II. Note: The complete 13 site Avr II restriction map of the E.coli genome has been reported. Methylation sensitivity The enzyme is not known to be affected by methylation. |
| Other Notes: | For life science research only. Not for use in diagnostic procedures. |
| Quality: | Absence of nonspecific endonuclease activities 1 μg λDNA is incubated for 16 hours in 50 μl SuRE/Cut Buffer H with an excess of Bln I. The number of enzyme units which do not change the enzyme-specific pattern is stated in the certificate of analysis. Absence of exonuclease activity Approximately 5 μg [3H] labeled calf thymus DNA are incubated with 3 μl Bln I for 4 hours at +37°C in a total volume of 100 μl 50 mM Tris-HCl, 10 mM MgCl2, 1 mM Dithioerythritol, pH approximately 7.5. Under these conditions, no release of radioactivity is detectable, as stated in the certificate of analysis. Typical ligation and recutting assay Bln I fragments obtained by complete digestion of 1 μg λ × EcoR I DNA ligated for 16 hours at +4°C with 1 U T4 DNA Ligase in 10 μl buffer that contains 66 mM Tris-HCl, 5 mM MgCl2, 5 mM Dithiothreitol, 1 mM ATP, pH 7.5 (at +20°C). The percentages of product that can be ligated and subsequently recut with Bln I and EcoR I (yielding the typical pattern of λ × EcoR I × Bln I fragments) are stated under "Lig" and "Rec" in the certificate of analysis. |
| Specificity: | Recognition sites: CCTAGG CCTAGG Restriction site: C↓CTAGG C↓CTAGG Heat inactivation: No inactivation of Bln I after incubation at 65 °C for 15 minutes. |
| Storage and Stability: | Do not store below −25°C. |
| Unit Definition: | One unit is the enzyme activity that completely cleaves 1 μg λ x EcoR I DNA fragments in one hour at +37 °C in a total volume of 25 μl (1x) SuRE/Cut Buffer H. |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| Flash Point(F) | does not flash |
| Flash Point(C) | does not flash |
| activity | specific activity: 10000 U/mL |
| Storage Temp. | −20°C |
| UNSPSC | 12352204 |
| Components | Enzyme Solution; SuRE/Cut Buffer H 10x concentrated |