Zero-background blunt-end topoisomerase cloning kit for PCR products from high-fidelity polymerases (KOD, Pfu, Phusion) or end-polished DNA, no ligase required. Standard reaction: 5 min at 25–37 °C; long or difficult inserts may need 10–15 min at 37 °C. Up to 10 kb is the manual’s stated design limit.
Key Features
- Blunt-end cloning for high-fidelity polymerase products
- 5-minute reaction at 25–37 °C (10–15 min at 37 °C for long or difficult inserts)
- Zero-background vector system
- Stated design limit: fragments up to 10 kb
Specifications
| Pack sizes | 20 × 10 µl or 40 × 5 µl (CYCV1601) / 80 × 10 µl or 160 × 5 µl (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?
The manual gives recommended DNA amounts for inserts from 100 bp to 5 kb and states a design limit of up to 10 kb. Reaction time is 5 minutes at 25–37 °C (10–15 minutes at 37 °C for long or difficult inserts); recombinants are selected by zero-background — no blue-white screening, no numeric positive rate stated.
Performance at a glance
| Insert size | Up to 10 kb (design limit); amounts given for 100 bp – 5 kb (10–180 ng) |
| Insert amount guide | 100–1000 bp → 10–40 ng; 1000–2000 bp → 40–80 ng; 2000–5000 bp → 80–180 ng |
| Reaction | 5 minutes at 25–37°C (37°C recommended), 10 µl system; 10–15 minutes at 37°C for long or difficult inserts |
| Selection | Zero background — no blue-white screening needed |
| Vector | 1865 bp, pUC ori, AmpR |
| Flanking restriction sites | Pst I, Hind III, EcoR V, EcoR I, Not I |
| Preferred insert ends | Blunt ends from high-fidelity polymerases (KOD, Phusion, Pfu); must NOT be 5′-phosphorylated |
| Storage | −20°C, stable 12 months |
| Pack sizes | 20 × 10 µl or 40 × 5 µl (CYCV1601) / 80 × 10 µl or 160 × 5 µl (CYCV1602) |
All values from the CYCV16 manual (PDF).
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.
- Set up the reaction at room temperature, not on ice; incubate 5 minutes at 25–37°C (10–15 minutes at 37°C for long inserts). Mix competent cells gently — do not pipette up and down.
Who sets these specifications
Changyu Bio supplies 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.
International ordering — CYCV16
- Single-unit orders are accepted, subject to confirmation — availability and lead time are confirmed on the quotation.
- Pack sizes: CV1601 = 40 reactions and CV1602 = 160 reactions in the 5 µl reaction format.
- International delivery, including Europe, the United States and Canada, Brazil and Latin America, the Middle East, Africa and Asia; destination and import requirements are confirmed before shipment; transit time and cold-chain packing for your destination are confirmed on the quotation.
- Shipped with ice packs; store at −20 °C on arrival.
- Payment by bank transfer (T/T) in US dollars after you confirm the quotation.
- Request a quotation for CYCV16 — tell us the quantity and destination city.
Alternative-product and purchasing questions
Is CYCV16 a blunt-end cloning alternative to Zero Blunt TOPO?
CYCV16 pTOPO-Blunt is an option for cloning blunt-ended PCR fragments, including products from proofreading polymerases. Review the CYCV16 vector map and protocol; the selectable markers and vector backbone need not match a Zero Blunt kit.
Can I clone A-tailed Taq products directly with CYCV16?
CYCV16 is a blunt-end route. Do not treat A-tailed and blunt-ended inserts as interchangeable: choose a TA kit for an appropriate A-tailed product or prepare the ends according to the relevant manual.
How do I request a quotation for CYCV16?
Include the current kit name or catalog number, requested quantity, delivery country and city, and the application details above. Ask us to confirm the available pack size, price and delivery conditions in writing. Tell us the insert-end type and vector features you require.
For laboratory research use only. Other brands identify the workflow being considered; no affiliation or endorsement is implied.