Hi, so i was looking for a expression plasmid with XbaI and HindIII restriction sites for 1908 bp insert. is there any sofrware to search the plasmid by restriction site? i've been looking for the compatible plasmid but after looking around 20 plasmid, i havent seen one with the right RES. Thankyou
Search on addgene database. It should be easy to find one plasmid if you know how to do it and what is correct source.
Snapgene is the software which can help you visualize.
You can use Plasmid: pSL1180 which has lots of multiple cloning sits inducing XbaI and HindIII
If you prepared your insert with PCR you can clone it with many other enzyme restriction sites in addition to XbaI and HindIII, you can even create hybrid sites that don't cut later. There are options like the ones above, or taking advantage of the flexibility of PCR.
QUOTE
"XbaI is blocked by overlapping dam methylation. If the XbaI recognition site is preceded by GA or followed by TC, the Dam methylase within most E. coli cloning strains will methylate the GATC site. For example, XbaI sites gaTˆCTAGA and TˆCTAGAtc will be blocked by methylation" https://international.neb.com/faqs/0001/01/01/is-xbai-affected-by-methylation
XbaI compatible overhangs: (T/CTAGA) -> AvrII, NheI, SpeI, StyI (C/CTAGG)
For HindIII there are no compatible overhangs.
Hi, thank you for the amazing answer. in the end i choose to add the restriction site at the primer that i use for the target gene amplification so that i can fit the target inside the general expression vector (i choose to use pET 21b+ and EcoRI, HindIII) Oscar L. Sierra Sepideh Fereshteh Abhijeet Singh Thank you very much
- Badges
- Science topic