Modern cloning methods which do not use restriction enzymes

LIC cloning 

Allows the creation of a plasmid from a single T4 DNA polymerase reaction from a plasmid and PCR reaction product. 

http://strucbio.biologie.uni-konstanz.de/ccp4wiki/index.php/LIC_cloning

SLIC, Gibson and CPEC cloning methods 

http://j5.jbei.org/j5manual/pages/22.html

Gateway Cloning

creates multiple plasmids from a single template plasmid. 

Yeast vectors, Protocols

We have 3 vectors in house at this time - Thank to Paul Jaschke and the Endy Lab:

• pRS315

http://www.ncbi.nlm.nih.gov/nuccore/U03441.1

• pRS315 with a bidirectional GAL promotor: 

http://www.ncbi.nlm.nih.gov/nuccore/K02115.1

NatureBioTechnology Reference

The pRS315 (ref. 32) vector had previously been modified by the insertion of a bidirectional gal1/10 promoter³³ between XhoI and HindIII sites. 

• pRS425

Here are a couple of protocol manuals for transforming yeast strain (W303) we have in house (which is His, Leu incapacitated)

Clontech Yeast Protocols