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Spe I
产品性质
| Quality Level【质量水平】 | 100 |
| biological source【生物来源】 | bacterial (Sphaerotilus spp.) |
| form【形式】 | solution |
| packaging【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 |
| parameter【参数】 | 37 ℃ optimum reaction temp. |
| shipped in【运输】 | dry ice |
| storage temp.【储存温度】 | −20℃ |
基本信息
| General description【一般描述】 | Spe I recognizes the sequence *A↓*CTAGT and generates fragments with 5′-cohesive termini. |
| Specificity【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). |
| Quality【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 Profile【DNA图谱分析】 | Number of cleavage sites on different DNAs
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| Unit Definition【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. |
| Analysis Note【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
Activity in PCR buffer: Not tested |
| Other Notes【其他说明】 | 仅用于生命科学研究。不可用于诊断。 |
| Components【组分】 | 组份不可单独销售 Enzyme Solution SuRE/Cut Buffer H 10x concentrated |
安全信息
| Storage Class Code【储存分类代码】 | 12 - Non Combustible Liquids |
| WGK | WGK 1 |
