Clontech's convenient "Mate & Plate” Libraries eliminate the time-consuming and labor-intensive cloning and amplification steps required in traditional two-hybrid library manufacturing and screening. These ready-to-go libraries require only simple co-culturing of the MATalpha library strain with your bait-expressing reporter strain (MATa), followed by selection on appropriate minimal medium.
Do it Yourself, Simply and Quickly
If our selection of ready-made libraries does not suit your needs, you may wish to Make Your Own Mate & Plate Library just the way we do it. Our system provides the materials and methods you need to create enough library vials for hundreds of yeast two hybrid screens—in less than a week.
Library creation occurs directly in our Y187 library yeast stain, utilizing the highly efficient homologous recombination machinery of Saccharomyces cerevisiae. There is no need for the labor-intensive library cloning, amplification, and harvesting in E. coli that are required by traditional methods of library construction.
The system uses SMART cDNA synthesis technology, which allows you to construct cDNA libraries from any tissue source starting with as little as 100 ng of total RNA.
What is SMART Technology?
Clontech’s SMART technology is based on two specific features of Moloney murine leukemia virus reverse transcriptase (MMLV RT):
- Terminal transferase activity
- Template switching activity
First-strand cDNA synthesis is primed by a modified oligo(dT) primer or random primers. When the MMLV RT reaches the 5’ end of the mRNA, the enzyme’s terminal transferase activity attaches non-template-directed nucleotides onto the newly synthesized strand of cDNA. Then the chemically modified SMART oligo pairs with the extended tail, and serves as a second template onto which the RT enzyme switches to complete first-strand synthesis.
SMART cDNA synthesis ultimately results in cDNA that contains known universal primer binding sequences at either end. As a result, SMART first-strand cDNA is:
- Available for PCR amplification, enabling you to start from nanogram amounts of RNA, so you can make a library from microdissected tissues, laser-captured cells, biopsy samples, etc.
- Homologous to the ends of the Matchmaker Gold prey plasmid, pGADT7-Rec; the library is created by cotransforming the yeast strain Y187 with pGADT7-Rec and the SMART cDNA.