Zero-background TA topoisomerase cloning kit for fast cloning of DNA fragments up to 10 kb generated by Taq DNA polymerase (3′-A overhang). Uses Vaccinia virus topoisomerase I strand-transfer activity for 5-minute ligation.
Key Features
- 5-minute room-temperature ligation (topoisomerase I)
- Zero-background vector system
- Clones fragments up to 10 kb with 3′-A overhangs
- Includes 1000 bp control insert for verification
Specifications
| Pack sizes | 20 × 10 µl or 40 × 5 µl (CYCV1401) / 80 × 10 µl or 160 × 5 µl (CYCV1402) |
| Components | pTOPO-TA 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
- TA cloning of Taq PCR products with 3′-A overhangs, up to 10 kb.
- 5-minute room-temperature ligation; 1000 bp control insert is in the box.
- DNA must not be 5′-phosphorylated. Competent cells are not included.
pTOPO-TA FAQ
Can I clone proofreading (blunt-end) PCR products with the TA kit?
Usually no. High-fidelity / proofreading polymerases generate blunt-ended products without 3′ A-overhangs, so TA ligation needs an A-tail first. The manual instructs end polishing/tailoring the PCR product with Taq DNA polymerase before the TA reaction. Alternatively, skip the A-tailing step entirely and use our pTOPO-Blunt Kit (CYCV16), which clones blunt products directly.
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 25–37 °C (10–15 minutes at 37 °C for long or difficult inserts).
How are recombinants selected?
The vector uses zero-background selection — no blue-white screening is required. The manual reports that with a 5 kb insert, at least 8 of 10 picked colonies contained the insert.
Are competent cells included?
No — the kit contains the pTOPO-TA vector and reaction components only; competent cells and ligase are not included. You transform with your own cells.
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, high-copy, pUC ori, AmpR |
| Flanking restriction sites | Pst I, Hind III, EcoR V, EcoR I, Not I |
| Preferred insert ends | 3′-A overhang from Taq PCR products; must NOT be 5′-phosphorylated |
| Storage | −20°C, stable 12 months |
| Pack sizes | 20 × 10 µl or 40 × 5 µl (CYCV1401) / 80 × 10 µl or 160 × 5 µl (CYCV1402) |
All values from the CYCV14 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. TA (A-overhang) ends from Taq 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# CYCV14 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# CYCV14 — 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 — CYCV14
- Single-unit orders are accepted, subject to confirmation — availability and lead time are confirmed on the quotation.
- 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, and air freight typically takes 7–14 days after dispatch.
- 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 CYCV14 — tell us the quantity and destination city.
Alternative-product and purchasing questions
Is CYCV14 an alternative to Invitrogen TOPO TA cloning kits?
CYCV14 pTOPO-TA is an option for cloning PCR products with single 3′-A overhangs. It is a TA-cloning workflow choice, not the same vector or a guarantee of identical clone recovery.
Which PCR products should I use with CYCV14?
Use the insert-end requirements in the CYCV14 manual. Taq-generated A-tailed products match the TA route; blunt products from proofreading polymerases require appropriate end treatment or a pTOPO-Blunt kit. Check the vector map and downstream restriction sites.
How do I request a quotation for CYCV14?
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.