Takara Yeast one-hybrid system—Matchmaker Gold

Overview

• Highest-performing yeast one-hybrid system
• Aureobasidin A selection eliminates screening background
• Construct and screen SMART cDNA libraries directly in yeast

Applications

• Yeast one-hybrid screening

Get results fast!

With the Matchmaker Gold system, one-hybrid library screening is straightforward, quick, and easy:

1. Create a bait construct by cloning 1–3 tandem repeats of the target DNA-binding sequence into pAbAi.
2. Create a bait-specific reporter strain by transforming and integrating the linearized pBait-AbAi construct into the Y1H Gold yeast strain and selecting on SD/–Ura agar medium.
3. Confirm integration of the bait sequence using colony PCR and the Matchmaker Insert Check PCR Mix 1.
4. Use SMART technology to synthesize cDNA containing ends homologous to the linearized pGADT7-Rec vector.
5. Create and screen your library in a single step by cotransforming your bait-specific reporter strain with SMART-generated cDNA and the linearized pGADT7-Rec vector, plating on AbA-containing selective medium.
6. Harvest colonies containing putative DNA-binding prey proteins and analyze library inserts using Matchmaker Insert Check PCR Mix 2.

Analysis by colony PCR

Screening positive colonies by PCR is a fast and convenient way to analyze the bait strain and sort through positive clones identified in Y1H and Y2H screens.

• Use Matchmaker Insert Check PCR Mix 1 to verify integration of the Y1H pBait-AbAi construct.
• Use Matchmaker Insert Check PCR Mix 2 to analyze inserts in positive clones from one-hybrid or two-hybrid screens.

Yeast one-hybrid media

Our yeast one-hybrid media sets contain a complete set of all the yeast media pouches needed for Matchmaker Gold One-Hybrid protocols.
For details, refer to the yeast one-hybrid media section on the Takara website.

Additional product information

Please see the product’s Certificate of Analysis for details on storage conditions, product components, and technical specifications.
Refer to the Kit Components List for information on included items.
Both documents are available under the Documents tab.