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 Clontech-SMART 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.
Yeast two-hybrid library construction process of clontech SMART system: Using total RNA as template, reverse transcribed into cDNA by SMART method, the amount of materials is small; Using high temperature resistant (60℃) reverse transcriptase, it can open more mRNA High-grade hairpin structure, the proportion of full length obtained is relatively high; Construction of a library can be used for yeast single-, two-, and three-hybrid;Library index is high.The library capacity of the original library is greater than 1×107 CFU, and the average inserted fragment is greater than 1000bp, the positive clone rate is greater than 95%.
Service introduction
Creative BioMart combines years of library construction experience to provide yeast two-hybrid library construction services in nuclear system based on the clontech-SMART method. Relying on the advantages of clontech's SMART method library construction system, it continues to optimize conditions to achieve a higher level of yeast nucleoprotein two-hybrid library construction.
Experiment workflow
- RNA extraction and single-stranded cDNA synthesis (SMART technology)
- Synthesis of double-stranded cDNA (LD-PCR) and cDNA separated by length
- Library gene recombination. Recombinant connection of cDNA and pGADT7 vector.
- Bacterial culture and quality inspection of the library
- Extract the library plasmid and transform it into yeast
a) The ligation product is electroporated into DH10b competent cells
Material requirements
- Total RNA > 20 μg
- Animal sample > 0.5 g
- Plant sample > 1 g
- Cell sample > 1*107
Delivery result
- Original nuclear library mother liquor
- Library plasmid
- Yeast library working solution
- Detailed experimental report, including complete experimental procedures and library detection results
Project cycle
It takes about 35 working days to complete nuclear system yeast two-hybrid library construction process based on clontech-SMART 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.