**** NEW: Relaxed PAM (TNTN) ttLbCas12a (pICSL90029) and multi-intron PAM-less Cas9 (pICSL90030) constructs have now been added to the Synbio repertoire. Also RUBY (He et al) non-invasive gene monitoring reporter (pICSL80094) construct added. ****
When using TypeIIS-mediated assembly we subscribe to “Standards for plant synthetic biology: A common syntax for exchange of DNA parts”, Patron et al (2015).
This page hosts a short course on Type IIS mediated cloning aimed at those that already have some basic knowledge of molecular cloning using standard restriction enzymes. The videos cover the background required to start using this powerful technique, including:
How typeIIS restriction enzymes enable the one-pot reaction
How to easily and reliably assemble multiple standard parts in a single reaction, and
How to make new parts
A lab protocol for the ‘One-Step, One-Pot Digestion-Ligation Reaction’ can be found HERE.
You can also download quick guides that include a re-cap of the background, how to design primers an laboratory protocols as two PDFs: 1 and 2. (NOTE –When printing these sheets, please select A3 paper, landscape, fitted to page.) If you are at TSL, you can also come and talk to us at any time for one-on-one help and training.
To obtain plasmids please visit the Plasmid Database and follow the instructions there. If you are at JIC/TSL, you will almost always be able to get DNA of any plasmid that you need within a day or two. If you are offsite you will need to obtain plasmids from Addgene.