Services
- Yeast Hybrid Services
- Yeast One-Hybrid (Y1H) System Service
- Yeast One-Hybrid (Y1H) Library Construction Service
- Yeast One-Hybrid (Y1H) Library Screening Service
- Prokaryotic Yeast One-Hybrid (Y1H) Service
- Genomic Yeast One-Hybrid (Y1H) Service
- Yeast One-Hybrid (Y1H) Point-to-Point Verification Service
- Yeast Library Construction Service for Transcription Factors
- Yeast Library Screening Service for Transcription Factors
- Transcription Factor Self-Activation Verification Service
- Yeast Two-Hybrid (Y2H) System Service
- Nuclear System Y2H Library Construction Service with Invitrogen Gateway Method
- Nuclear System Y2H Library Construction Service with Clontech-SMART Method
- Membrane System Y2H Library Construction Service with Invitrogen-Gateway Method
- Membrane System Y2H Library Construction Service with Clontech-SMART Method
- Reverse Yeast Two-Hybrid (rY2H) Library Construction Service
- Yeast Two-Hybrid (Y2H) Library Screening Service
- Nuclear System Yeast Two-Hybrid (Y2H) Library Screening Service
- Membrane System Yeast Two-Hybrid (Y2H) Library Screening Service
- Nuclear System Yeast Two-Hybrid (Y2H) Verification Service
- Membrane System Yeast Two-Hybrid (Y2H) Verification Service
- High-throughput Yeast Two-Hybrid (Y2H) Library Screening Service
- Prokaryotic Yeast Two-Hybrid (Y2H) Service
- Genomic Yeast Two-Hybrid (Y2H) Service
- Mammalian Two-Hybrid (mY2H) Service
- Yeast One-to-One Verification Service
- Yeast Three-Hybrid (Y3H) System Service
- Yeast One-Hybrid (Y1H) System Service
- Yeast Expression System Services
- Yeast Foreign Gene Secretion and Expression Service
- Yeast Signal Peptide Tracking and Screening Service
- Yeast Stress Resistance Gene Screening Service
- Yeast Gene Knockout Service
- Yeast Surface Display Service
- Yeast Display Service of IgG Full-length Antibody
- Pichia pastoris Protein Expression Analysis
- Saccharomyces cerevisiae Protein Expression Analysis
- DNA Shuffling
- Yeast RNAi System
- Yeast Detection Services for Protein Functional Domain
- Yeast Microarray Chip Service
Online Inquiry
Nuclear System Y2H Library Construction Service with Invitrogen Gateway Method
The nuclear system yeast two-hybrid (Y2H) technology is based on the transcription activation domain (AD) and DNA binding domain (BD) of transcription factors to construct fusion genes respectively, and detect protein interactions in the nucleus by activating the expression of reporter genes. After continuous optimization and improvement, it can not only detect interactions between known proteins, but also discover unknown proteins that interact with known proteins. It has been widely used in the fields of proteomics, cell signal transduction and functional genomics, and has become an important experimental method in the field of molecular biology research.
Gateway system uses site-specific recombination technology (Cloneminer cDNA library construction kit) to construct a library. It reverse-transcribes mRNA into cDNA to construct a library, without PCR, to maximize the quality and fidelity of the library. Using gateway's high-efficiency fragment and vector recombination ligation method has high ligation efficiency, which can greatly increase the original library capacity (original library capacity above 1*107 CFU).
Service introduction
Creative BioMart combines years of library construction experience to provide yeast two-hybrid library construction services based on the Gateway method. Relying on the advantages of invitrogen's gateway method library construction system, it continues to optimize conditions to achieve a higher level of yeast nucleoprotein two-hybrid library construction.
Experiment workflow
Total RNA extraction and mRNA isolation
cDNA synthesis (direct synthesis using enzymatic method, without PCR amplification)
cDNA length classification
- 1
Use the gateway BP method to recombine the graded cDNA and pDONR222 vector to form a primary library
- 2
Electrotransformation and plating culture of primary library
Primary library detection
Positive primary library plating amplification and plasmid extraction
- 3
Use the gateway LR method to recombine the primary library plasmid and the yeast library vector pGADT7-GW to form a secondary library
- 4
Electrotransformation and plating culture of secondary library
Secondary library detection
Positive secondary library plating amplification and plasmid extraction
- 5
Material requirements
- Total RNA > 600 μg
- Animal sample > 1 g
- Plant sample > 6 g
- Cell sample > 1*107
Delivery result
- Mother liquor (LB medium containing glycerol) and plasmids of the primary library
- Mother liquor (LB medium containing glycerol) and plasmids of the secondary library
- Construction report (electronic version), including detailed experimental procedures and library construction picture (JPG).
Project cycle
It takes about 45 working days to complete nuclear system yeast two-hybrid library construction process based on invitrogen-gateway method.
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.