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| Product Name | Homing Endonuclease I-SceI (Recognition Site: 5'-TAGGGATAA↓CAGGGTAAT-3') |
| Catalog No. | IVDRM-0167 |
| Description | I-SceI is an intron-encoded homing endonuclease derived from Saccharomyces cerevisiae. It specifically recognizes and cleaves an 18 bp non-palindromic sequence (5'-TAGGGATAACAGGGTAAT-3'), generating 4 bp 3' overhang sticky ends. Because its recognition sequence is exceptionally long, it occurs in genomes with extremely low probability—approximately once per 7×10^10 random bases—making I-SceI an ideal tool for precise genome manipulation. |
| Intended Use | Precise genome engineering and large-scale (whole-genome level) targeted cleavage; introduction of site-specific double-strand breaks at engineered I-SceI sites for genome manipulation workflows. |
| Principle / Technology | Group II intron-encoded homing endonuclease (meganuclease) that recognizes an 18 bp non-palindromic sequence and cleaves it to generate 4 bp 3' overhangs. The exceptionally long recognition site occurs naturally with a probability of only about once per 7×10^10 random bases, enabling highly specific genome-scale cleavage. |
| Sample Type | Double-stranded DNA substrates such as pUC-I-SceI; genomic DNA containing engineered I-SceI recognition sites. |
| Performance Range / Specifications | Concentration: 5 U/μL Recognizes an 18 bp non-palindromic sequence; generates 4 bp 3' overhang sticky ends Natural occurrence probability of the site: ~1 per 7×10^10 random bases |
| Specificity | Specifically recognizes and cleaves 5'-TAGGGATAA↓CAGGGTAAT-3' / 3'-ATCCC↑TATTGTCCCATTA-5', producing 4 bp 3' overhangs. |
| Reaction Conditions / Protocol | 1× universal reaction buffer; incubate at 37°C. Heat inactivation: 80°C for 20 min. |
| Components / Formulation | I-SceI homing endonuclease (5 U/μL); universal reaction buffer. |
| Storage Conditions | Store at -25 to -15°C. |
| Shelf Life | 2 years when stored at -25 to -15°C. |
| Package Specifications | 250 U |
| Concentration | 5 U/μL |
| Activity / Unit Definition | One unit (U) is defined as the amount of enzyme required to digest 1 μg of pUC-I-SceI DNA in a reaction at 37°C for 1 h. |
| Source / Origin | Saccharomyces cerevisiae (group II intron-encoded endonuclease). |
| Expiration Date / Stability | Stable for 2 years at -25 to -15°C. |
| Recommended Buffer System | 1× universal reaction buffer. |
| Application Notes / Precautions | The 18 bp recognition sequence occurs at extremely low frequency (~1 in 7×10^10 random bases), making I-SceI well suited for precise, genome-scale cleavage. Cleavage generates 4 bp 3' overhang sticky ends. |
| Recognition Site | 5'-TAGGGATAA↓CAGGGTAAT-3' 3'-ATCCC↑TATTGTCCCATTA-5' |
| Heat Inactivation | 80°C for 20 min. |
| Methylation Sensitivity | No effect from methylation. |
For research use only, not for clinical use.
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