The pLPS-B blunt topo vector is a high efficient and very convenient tool for a direct cloning of PCR products. The type I topoisomerase from Vaccinia virus can cut and rejoin the DNA fragment without the action of DNA ligase.Topoisomerase I premixed in the vector solution rejoins the input DNA fragments to vector ends within 5 min at 37℃. There is no need to consider whether insert DNA has been phosphorylated. Furthermore, there is no need to use IPTG/X-gal in the transformation process due to its high ligation and cloning efficiency (up to 90%). This means you can obtain 9 real clones from 10 randomly picked transformed colonies.The pLPS-B blunt vector is suitable for the cloning of blunt ended DNA fragments. If your DNA has 3'-A tails, you should remove A tails by appropriate methods before start.In general, we recommend the use of high fidelity DNA polymerases, like Pfu, Pfu plus, PyroMax, to produce the blunt ended PCR products. By using pLPS-B vector system, up to 10 kbp sized DNA fragments can be easily cloned without the remarkable loss of efficiency.
The pLPS-B blunt vector is provided in 50% buffered glycerol. Simply mix 1 ㎕ of vector solution with 1 ㎕ of 5x reaction buffer and your DNA to final 5 ㎕ volume, and incubate for 5 min at 37℃. Then you are already ready to transform E.coli. In the next day, you can get the clone in plasmid.
- 25 ㎕ pLPS-B Blunt Vector- 50 ㎕ 5x Topo Reaction Buffer- 5 ㎕ Control Insert DNA (blunt ended)
Kit Component : EBK-1001 QC tests- Performance tests. Storage Condition
- Store at -20℃.