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| Product Name | Restriction Enzyme ApeKI (Recognition Site: 5'-G↓CWGC-3') |
| Catalog No. | IVDRM-0149 |
| Description | ApeKI is a Type IIP restriction endonuclease originally isolated from the hyperthermophilic aerobic archaeon Aeropyrum pernix K1. It recognizes and cuts the degenerate sequence G/CWGC, generating cohesive ends with a 3-nucleotide overhang. ApeKI is one of the enzymes of choice for genotyping-by-sequencing (GBS) and is also widely used in animal and plant genetics research, population evolution analysis, and functional gene mapping. |
| Intended Use | Genotyping-by-sequencing (GBS) library preparation, as well as animal and plant genetics research, population evolution analysis, and functional gene mapping. |
| Principle / Technology | ApeKI is a Type IIP restriction endonuclease that recognizes the degenerate sequence 5'-G↓CWGC-3' (W = A or T) and cleaves within the recognition site, producing sticky ends with a 3-nucleotide overhang. Originating from a hyperthermophilic archaeon, the enzyme is highly thermostable and operates at an optimal reaction temperature of 75°C. |
| Sample Type | DNA substrates such as lambda DNA (λDNA) bearing ApeKI recognition sites. |
| Performance Range / Specifications | Concentration: 10 U/μL. Unit definition: One unit (U) is the amount of enzyme required to completely digest 1 μg of λDNA in 1 hour at 75°C in a 50 μL reaction. Cleavage generates cohesive ends with a 3-nucleotide overhang. CpG methylation of the recognition site blocks cleavage. |
| Specificity | Specifically recognizes and cuts 5'-G↓CWGC-3' (W = A or T; complementary strand 3'-CGWC↑G-5'). |
| Reaction Conditions / Protocol | Optimal reaction: incubate at 75°C in 1× Cut Buffer C. ApeKI cannot be heat-inactivated; to terminate the reaction, remove the enzyme by phenol-chloroform extraction or column purification. |
| Components / Formulation | ApeKI restriction enzyme (10 U/μL) and 1× Cut Buffer C. |
| Storage Conditions | Store at -25°C to -15°C. |
| Shelf Life | Valid for 2 years when stored at -25°C to -15°C. |
| Package Specifications | 500 U |
| Concentration | 10 U/μL |
| Activity / Unit Definition | One unit (U) is defined as the amount of enzyme required to completely digest 1 μg of λDNA in 1 hour at 75°C in a 50 μL reaction system. |
| Source / Origin | Derived from the hyperthermophilic aerobic archaeon Aeropyrum pernix K1. |
| Expiration Date / Stability | Stable for 2 years from the date of manufacture when stored at -25°C to -15°C. |
| Recommended Buffer System | 1× Cut Buffer C (supplied). |
| Application Notes / Precautions | ApeKI is highly thermostable and cannot be heat-inactivated. Terminate reactions by phenol-chloroform extraction or column purification before downstream steps. CpG methylation of the recognition site blocks cleavage, which is exploited in methylation-sensitive GBS applications. |
| Recognition Site | 5'-G↓CWGC-3' 3'-CGWC↑G-5' |
| Heat Inactivation | Cannot be heat-inactivated; use phenol-chloroform extraction or column purification to remove the enzyme. |
| Methylation Sensitivity | CpG methylation: cleavage is blocked. |
For research use only, not for clinical use.
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