HE Swift Cloning Kit
TOOLS HE Swift Cloning Kit is an advanced positive selection system for the highest efficiency cloning of PCR products generated with Pfu DNA polymerase, Taq DNA polymerase, or other thermo-stable DNA polymerases. Additionally, any other DNA fragment, either blunt or sticky-end, can be successfully cloned using the kit. Cloning is fast and efficient; ligation takes only 5 minutes and yields more than 99% positive clones.
The kit features the novel positive selection cloning vector pHE. This vector contains a lethal gene which is disrupted by ligation of a DNA insert into the cloning site. As a result, only cells with recombinant plasmids are able to propagate, eliminating the need for expensive blue/white screening.
The vector contains an expanded multiple cloning site, as well as a T7 promoter for in vitro transcription. Sequencing primers are included for convenient sequencing of the insert.
Quick and efficient: Blue/white screening is not necessary, and the whole ligation process can be finished within 5 min.
Sensitive and widely applied: Good for the ligation of low concentration fragments and long fragments, works for both blunt and sticky end product.
No need for blue white screening.
Ligation time: 5 minutes.
Suitable for blunt-end and sticy-end PCR product.
pHE Vector Map:
Data 1: Insert PCR Product from HE Swift Cloning KitFig 1. Insert product (size: 909 bp).M, Marker; Lane 1, Insert PCR product.
Data 2: Ligation Mixture PCR from HE Swift Cloning KitFig 2. Ligation mixtureM, Marker; Lane 1, ligation mixture
Data 3: Colony PCR determining positive clone from HE Swift Cloning KitFig 3. Colony PCR
WU, Tsung-Ru, et al. Gut commensal Parabacteroides goldsteinii plays a predominant role in the anti-obesity effects of polysaccharides isolated from Hirsutella sinensis. Gut, 2018, gutjnl-2017-315458.
PENG, Shih‐Yuan, et al. miR‐134 targets PDCD7 to reduce E‐cadherin expression and enhance oral cancer progression. International journal of cancer, 2018, 143.11: 2892-2904.
Q1.HEswift cloning kit有推薦使用哪種Competent cell嗎?
Q2.HEswift cloning kit的效率不好，應該如何改善?
A:cloning的過程當中，有很多關鍵的點可以注意，其中insert和vector是否有經過PCR clean up或Gel elution的方式處哩，可提高ligation的效率，同時ligation時要注意insert/vector的比例，接著Transformation的時候，也要注意是否操作無誤。