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| Product Name | Rapid TA/Blunt Zero-Background Cloning Kit |
| Catalog No. | NATR-HMM-0169 |
| Description | A topoisomerase-based cloning kit that combines TA and blunt-end compatibility with lethal-gene counterselection to minimize empty-vector background. |
| Intended Use | TA cloning and blunt-end cloning of PCR products. |
| Principle / Technology | Insertion of the target fragment disrupts a lethal selection cassette in a dual-resistant cloning vector, enabling strong counterselection against nonrecombinant background. |
| Sample Type | A-tailed or blunt-ended PCR products. |
| Performance Range / Specifications | Five-minute reaction at room temperature; reported positive-clone rate above 95%. |
| Specificity | Lethal-gene counterselection suppresses colonies derived from nonrecombinant vector. |
| Reaction Conditions / Protocol | Add the DNA fragment to the cloning mixture and incubate for 5 minutes at room temperature before transformation. |
| Components / Formulation | The 25-reaction format contains 25 µL of 5× TA/blunt zero-background cloning mix, 5 µL of a 500 bp control insert at 20 ng/µL, and 200 µL of a 10 µM M13 forward/reverse primer mix. The 50-reaction format contains two 25 µL tubes of cloning mix, 10 µL of control insert, and 400 µL of primer mix. |
| Storage Conditions | Store at −30 to −15°C. |
| Package Specifications | 25 reactions; 50 reactions. |
| Product Form | Rapid TA/blunt cloning kit. |
| Quality Control | Cloning performance is assessed with control inserts and colony screening primers. |
| Key Features | TA and blunt-end compatibility; five-minute reaction; positive-clone rate above 95%; lethal-gene counterselection; dual antibiotic selection. |
| Shipping Conditions | Ship at −20 to 0°C. |
| Compatibility | Compatible with A-tailed and blunt-ended PCR products and standard cloning competent cells. |
| Application Notes / Precautions | For low-concentration inserts, concentrate the recovered DNA when it is below 20 ng/µL. If the insert is highly concentrated, dilute it so that at least 1 µL is added. When an unpurified PCR product is generated from a circular plasmid template, digest the parental template with DpnI or purify the target band before cloning. |
For research use only, not for clinical use.
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