CYCV2001 is a directional topoisomerase cloning kit that places a blunt-end PCR fragment, amplified with a high-fidelity polymerase, into a Gateway entry vector in one step at room temperature in 5 minutes. The entry clone can be moved into Gateway destination vectors by LR recombination to give expression clones for prokaryotic, mammalian, yeast or insect systems.
Catalog no. CYCV2001 · For research use only
Request a quote · Download manual (PDF)
Key features
- Directional TOPO cloning into a Gateway entry vector in 5 minutes at room temperature
- More than 90% of inserts in the correct orientation, reducing screening work
- Entry clones can be moved to Gateway destination vectors by LR recombination to switch expression systems
- The entry clone has no prokaryotic Shine-Dalgarno (SD) sequence; choose a destination vector with an SD sequence for E. coli expression
- Stored at -20 °C for 12 months; for research use only
Catalog numbers and pack sizes
| Catalog # | Product | Pack size |
|---|---|---|
| CYCV2001 | pTOPO-ENTR/D Directional Cloning Kit | 20 reactions |
Specifications
| Vector type | Gateway entry vector, directional TOPO |
|---|---|
| Insert | Blunt-end PCR product from a high-fidelity polymerase |
| Reaction | 5 min, room temperature |
| Orientation | >90% correct |
| Downstream | LR recombination with destination vectors |
| Storage | -20 °C, 12 months |
| Use | Research use only |
Applications
- Building Gateway entry clones
- Moving one gene between prokaryotic, mammalian, yeast and insect expression systems
- Directional cloning of blunt PCR products
Ordering
International ordering — CYCV2001
- Single-unit orders are accepted, subject to confirmation — availability and lead time are confirmed on the quotation.
- Different Changyu Bio and branded items can be combined into one shipment.
- 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.
- Storage: -20 °C, 12 months. Shipping temperature is confirmed per item.
- Payment by bank transfer (T/T) in US dollars after you confirm the quotation.
- Request a quotation for CYCV2001 — tell us the quantity and destination city.
pTOPO-ENTR/D Directional Cloning Kit FAQ
What insert does it need?
A blunt-end, non-phosphorylated PCR fragment amplified with a high-fidelity polymerase. Add CACC to the 5′ end of the forward primer; do not add CACC to the 5′ end of the reverse primer. Follow the manual for stop-codon design; clones are selected on kanamycin.
Can I use it for E. coli expression?
Yes, but the entry clone has no Shine-Dalgarno (SD) sequence, so choose a destination vector that carries one for the LR reaction.
How long does cloning take?
Five minutes at room temperature.
How do I order CYCV2001?
Request a quote with the catalog number, quantity and destination. Single-unit orders are accepted; availability, lead time and shipping conditions are confirmed on the quotation, payment is by USD bank transfer, and items can be combined into one shipment.
Browse related products: Ptopo cloning kits.