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

More Information

Applications

• Yeast one-hybrid screening

Get results fast!

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

Step 1: Create a bait construct by cloning 1–3 tandem repeats of the target DNA-binding sequence into pAbAi.

Step 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.

Step 3: Confirm the integration of the bait sequence using colony PCR and the Matchmaker Insert Check PCR Mix 1.

Step 4: Use SMART technology to synthesize cDNA containing ends that are homologous to the ends of the linearized pGADT7-Rec vector.

Step 5: Create and screen your library in a single step by cotransforming your bait-specific reporter strain with the SMART-generated cDNA and the linearized pGADT7-Rec vector, and plating on AbA-containing selective medium.

Step 6: Harvest the resulting colonies, which contain putative DNA-binding prey proteins, and analyze your library inserts using the 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 the positive clones identified in Y1H and Y2H screens. With the Matchmaker Insert Check PCR Mix 1, you can verify the integration of your Y1H pBait-AbAi construct, and with the companion Matchmaker Insert Check PCR Mix 2, you can quickly sort through and analyze the inserts in positive clones that have emerged from either one-hybrid or two-hybrid screens.

Yeast one-hybrid media

Our yeast one-hybrid media sets each contain a complete set of all the yeast media pouches you need for Matchmaker Gold One-Hybrid protocols.

Additional product information

Please see the product`s Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.