Zero-background blunt-end topoisomerase cloning kit for fragments up to 10 kb generated by high-fidelity polymerases (KOD, Pfu, Phusion) or end-polished DNA. Fast, reliable cloning without ligase.
Key Features
- Blunt-end cloning for high-fidelity polymerase products
- 5-minute topoisomerase ligation at room temperature
- Zero-background vector system
- Fragments up to 10 kb
Specifications
| Pack sizes | 20 rxns (CYCV1601) / 80 rxns (CYCV1602) |
| Components | pTOPO-Blunt Vector (30 ng/µl), 1000 bp Control, 10× Enhancer |
| Storage | -20°C; stable for 12 months |
| Shipping | Ice packs; store at the listed temperature on arrival |
Typical Applications
- Cloning blunt PCR products from high-fidelity polymerases, up to 10 kb.
- 5-minute ligation; 1000 bp control insert included.
- DNA must not be 5′-phosphorylated. Prefers blunt ends, not A-overhangs.
pTOPO-Blunt FAQ
Can I clone proofreading PCR products directly?
Yes — this kit is designed for blunt-ended inserts, so products from high-fidelity / proofreading polymerases (e.g., Pfu-type mixes such as our FastHiFi or LongHiFi) ligate directly with no A-tailing step.
Do I need an A-tailing step?
No. A-tailing is only needed when cloning blunt products into a TA vector — that workflow is covered by our pTOPO-TA Kit (CYCV14) after Taq end-polishing per its manual.
What insert sizes does the manual support?
Insert fragments from 100 bp to 5 kb are demonstrated in the manual, with a design limit of up to 10 kb. Reaction time is 5 minutes at room temperature; recombinants are selected by zero-background — no blue-white screening, no numeric positive rate stated.
Limits and handling
- For laboratory research use only. Not for diagnostic or therapeutic use.
- Shipped with ice packs. Store at the temperature in the specifications table on arrival.
- Store at −20°C (manual: 12 months).
- DNA fragments must not be 5′-phosphorylated. Blunt ends from high-fidelity enzymes (KOD, Pfu, Phusion) are preferred. Inserts up to 10 kb.
- Incubate the cloning reaction no more than 5 minutes at 15–30°C. Mix competent cells gently — do not pipette up and down.
Who sets these specifications
Changyu Bio manufactures Cat# CYCV16 and is responsible for the specifications on this page. Confirm pack sizes, storage, and protocol against the PDF manual before use. Quotes: market@changyubio.com or WhatsApp +86 18665577773.
Cat# CYCV16 — For research use only (RUO). Not for diagnostic or clinical use. Contact market@changyubio.com or WhatsApp +86 18665577773 for pricing and export delivery to Africa, Southeast Asia and worldwide.