产品说明
一般描述
Spe I recognizes the sequence *A↓*CTAGT and generates fragments with 5′-cohesive termini.
特异性
Recognition sites: *A*CTAGT
*A*CTAGT
Restriction site: *A↓*CTAGT
*A↓*CTAGT
Heat inactivation: Spe I can be heat inactivated by incubation at 65 ℃ for 15 minutes (up to 100 U/μg DNA).
质量
Absence of nonspecific endonuclease activities
1μg Ad2 DNA is incubated for 16 hours in 50μl SuRE/Cut Buffer H with an excess of Spe 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 5μg [3H] labeled calf thymus DNA are incubated with 3μl Spe I for 4 hours at +37℃ in a total volume of 100μl 50mM Tris-HCl, 10mM MgCl2, 1mM Dithioerythritol, pH approximately 7.5. Under these conditions, no release of radioactivity is detectable, as stated in the certificate of analysis.
DNA图谱分析
Number of cleavage sites on different DNAs
- λ: 0
- φX174: 0
- Ad2: 3
- M13mp7: 0
- pBR322: 0
- pBR328: 0
- pUC18: 0
- SV40: 0
单位定义
One unit is the enzyme activity that completely cleaves 1 μg Ad2 DNA in one hour at +37 ℃ in a total volume of 25 μl SuRE/Cut Buffer H.
分析说明
Compatible ends
Spe I ends are compatible with ends generated by Bln I, Nhe I, and Xba I.
Isoschizomers
The enzyme is an isoschizomer of Bcu I and Ahl I.
Methylation sensitivity
As indicated by (*) on the recognition sequence above, Spe I is inhibited by the presence of N6-methyladenine and 5′-methylcytosine ( mA↓m CTAGT).
Incubation temperature
+37℃
PFGE tested
Spe 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 10U of enzyme/μg DNA and 4 hour incubation time.
Ligation and recutting assay
Spe I fragments obtained by complete digestion of 1μg Ad2 DNA are ligated with 1U T4 DNA Ligase in a volume of 10μl by incubation for 16 hours at +4℃ in 66mM Tris-HCl, 5mM MgCl2, 5mM Dithiothreitol, 1mM ATP, pH 7.5 (at +20℃), resulting in >90% recovery of Ad2 DNA.
Subsequent re-cutting with Spe I yields >95% of the typical pattern of Ad2 × Spe I fragments.
SuRE/Cut Buffer System
The buffer in bold is recommended for optimal activity
- A: 75-100%
- B: 75-100%
- H: 100%
- L: 75-100%
- M: 100%
Activity in PCR buffer: Not tested
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 (Sphaerotilus spp.) |
| 形式 | solution |
| 包装 | pkg of 1,000 U (11008951001 [10 U/μl]) pkg of 1,000 U (11207644001 [40 U/μl]) pkg of 200 U (11008943001 [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 |
