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| Product Name | Restriction Enzyme SgeI (Recognition Site: 5'-m5CNNG(N)9↓-3') |
| Catalog No. | IVDRM-0158 |
| Description | SgeI is a methylation-dependent restriction endonuclease that cleaves DNA targets containing 5-methylcytosine in either single-stranded or double-stranded DNA. It recognizes the m5CNNG(9/13) site and delivers optimal cleavage at 37°C in its dedicated reaction buffer. To ensure consistent performance, the enzyme storage buffer contains premixed BSA, which enhances enzyme stability and binds potential contaminants that may be present in DNA preparations. |
| Intended Use | Site-specific cleavage of DNA containing 5-methylcytosine (m5C) in single- or double-stranded DNA substrates; suitable for molecular cloning and methylation-related molecular biology workflows. |
| Principle / Technology | Type II restriction endonuclease that recognizes the m5CNNG(9/13) sequence and cleaves double-stranded DNA at fixed positions downstream of the recognition site, with cleavage dependent on 5-methylcytosine modification. |
| Sample Type | Single-stranded or double-stranded DNA substrates containing 5-methylcytosine, e.g., pUC19-SgeI DNA (Dcm+). |
| Performance Range / Specifications | Concentration: 5 U/μL Optimal cleavage in 1×SgeI Buffer at 37°C Isoschizomers: Bse3DI, BseMI |
| Specificity | Recognizes and cleaves DNA containing 5-methylcytosine at the m5CNNG(9/13) site; consistently cleaves DNA methylated by Dcm methyltransferase and cuts targets overlapping CpG-methylated sequences. |
| Reaction Conditions / Protocol | 1×SgeI Buffer; incubate at 37°C. Heat inactivation: 80°C for 20 min. |
| Components / Formulation | SgeI restriction enzyme (5 U/μL) supplied in a storage buffer containing premixed BSA for enhanced stability; dedicated 1×SgeI 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, added incrementally to a 50 μL reaction containing 1 μg of pUC19-SgeI DNA (Dcm+) substrate incubated at 37°C for 1 h, at which the digestion product banding pattern no longer changes with further enzyme addition. |
| Expiration Date / Stability | Stable for 2 years at -25 to -15°C. |
| Compatibility | Isoschizomers: Bse3DI, BseMI. |
| Recommended Buffer System | 1×SgeI Buffer (dedicated reaction buffer). |
| Application Notes / Precautions | Premixed BSA in the storage buffer improves enzyme stability and sequesters potential contaminants in DNA preparations; use the dedicated SgeI buffer for optimal cleavage. |
| Recognition Site | 5'-m5CNNG(N)9↓-3' 3'-GNNC(N)13↑-5' |
| Heat Inactivation | 80°C for 20 min. |
| Methylation Sensitivity | Dcm: always cleaves DNA methylated by Dcm methyltransferase. CpG: cleaves targets overlapping CpG-methylated sequences. |
For research use only, not for clinical use.
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