产品说明
一般描述
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.
特异性
Recognition sites: CCTAGG
CCTAGG
Restriction site: C↓CTAGG
C↓CTAGG
Heat inactivation: No inactivation of Bln I after incubation at 65 ℃ for 15 minutes.
质量
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℃ 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℃ 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℃). 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.
DNA图谱分析
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
单位定义
One unit is the enzyme activity that completely cleaves 1 μg λ x EcoR I DNA fragments in one hour at +37 ℃ in a total volume of 25 μl (1x) SuRE/Cut Buffer H.
分析说明
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%
Activity in PCR buffer: Not tested
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.
其他说明
仅用于生命科学研究。不可用于诊断。
产品性质
| 质量水平 | 100 |
| 生物来源 | bacterial (Brevibacterium linens) |
| 形式 | solution |
| 包装 | pkg of 1,000 U (11558170001 [10 U/μl]) pkg of 200 U (11558161001 [10 U/μl]) |
| manufacturer/tradename | Roche |
| 参数 | 37 ℃ optimum reaction temp. |
| 运输 | dry ice |
| 储存温度 | −20℃ |
组份列表
组份不可单独销售
| 货号 | 组份 |
| Enzyme Solution | |
| SuRE/Cut Buffer H 10x concentrated |
安全信息
| 储存分类代码 | 12 - Non Combustible Liquids |
| WGK | WGK 1 |
| 闪点(F) | does not flash |
| 闪点(C) | does not flash |
