Yeast Three-Hybrid (Y3H) Library Screening Service

The yeast three-hybrid system is mainly used to study more complex macromolecular interactions including three components, providing a new method for the study of protein-protein, RNA-protein, and small molecule-protein interactions.

The yeast three-hybrid technology is improved from the yeast two-hybrid technology. This system introduces a pBrige plasmid, which is characterized by the ability to insert two foreign proteins at the same time. The N-terminus of the protein inserted by MCSII is still fused with the DNA binding domain (BD) in the Gal4 system, and no other tags are fused. But there is a Pmet25 promoter upstream of it. The feature of this promoter is that it can only work when methionine (Met) is lacking. Therefore, the experiment must be carried out in a methionine-deficient medium. The pBrige plasmid was also transformed into Y2HGold strain. At the same time, the third protein gene was cloned into pGADT7 vector, and then transformed into AH109 yeast strain. Then combine these two bacteria in a small amount, and the rest of the operation steps are the same as the yeast two-hybrid system. The only difference is that all defective media must be configured on the basis of methionine deficiency. If these three proteins can interact, they can grow on the five-deficient medium (SD/-Met/-Ade/-His/-Leu/-Trp).

Service introduction

Creative BioMart combines many years of experience in yeast related research to provide yeast three-hybrid library screening service, which has great advantages in technology and quality.

Experiment process

1. Construction of bait vector

2. Self-activation detection of bait plasmid

3. Co-transformation of competent yeast cells with library plasmids and bait plasmids

4. Library screening and 3AT optimization

5. Sequencing analysis of library screening results

6. Screening results rotation verification

7. Organize the data and make detailed experimental reports

Delivery result

• Detailed and complete experiment report
• Clear experiment pictures
• Sequencing results of screening results
• Plasmid

Project cycle

It takes about 20 working days to complete yeast three-hybrid (Y3H) library screening service.

Service process

Please let us know the details of your yeast related projects through online consultation. We will reply to you in time and provide you with a comprehensive evaluation report on your project, including the feasibility evaluation of the project and the detailed experimental plan designed by our professionals. As our condition optimization is completed, the result report and other final products will be delivered to you immediately.

Creative BioMart has accumulated many years of experience in the field of yeast research, helping our customers accelerate research in all fields using yeast as a research tool, and improve the overall success rate of the project. Please tell us your project requirements, and we will provide you with a full service from strategy design to final report. If you have any questions, please feel free to contact us.