Yeast One-Hybrid (Y1H) Detection Service

Yeast One-Hybrid

High-Throughput Screening

Low False Positive Rate

Service Features

Yeast One-Hybrid (Y1H) is an experimental technique used to study protein-DNA interactions. It combines transcriptional activation and detection systems within yeast cells to screen and identify proteins that interact with specific DNA sequences, offering advantages such as high-throughput screening, physiological relevance, low false positive rates, quantitative analysis capabilities, and applicability across multiple research fields.

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Service Introduction
Service Advantages
Service Process
Customer Requirements
Service Description
Case Studies
Q&A
Customer reviews

Service Introduction

Yeast One-Hybrid technology is a tool developed from the yeast two-hybrid system for studying nucleic acid-protein interactions. It is widely used to investigate gene expression regulation in eukaryotic cells, such as identifying DNA binding sites, discovering potential binding protein genes, and analyzing DNA binding domain information.

Service Advantages

One-stop auxiliary detection method for convenient and reliable experimental conclusions
Years of library construction experience with 4 optimized Total RNA extraction technical solutions to ensure Total RNA purity and integrity, maintaining sample diversity
Ability to study multiple protein interactions simultaneously, enabling high-throughput screening and analysis for rapid acquisition of extensive interaction information
Implementation of multiple screening steps, such as reporter gene validation and dual selection, to reduce false positives from non-specific interactions

Service Process

Yeast One-Hybrid Library Screening

Receive order and samples

Bait vector and bait strain construction

Auto-activation detection

Library screening

Positive clone strain identification

Result delivery

Yeast Hybrid Library Construction

Total RNA extraction

mRNA isolation and purification

Double-stranded cDNA synthesis and purification

Fusion of double-stranded cDNA with vector

E. coli library culture

Library plasmid

Yeast Hybrid Validation

Bait and prey recombinant plasmid construction

Bait auto-activation and toxicity validation

Yeast cell co-transformation validation

Dilution spot assay validation

note:Yeast One Hybrid.

Customer Requirements

1. Species information, sample type

2. Gene sequence information

3. Experimental purpose

Service Description

Customer Provides	Final Deliverables
Tissue/Cells	Yeast library culture with titer above 1×10⁶ cfu/mL; Library PCR identification result images (positive rate above 90%)
Promoter gene sequence/plasmid	All original images from the screening process; Sequencing results of all interacting prey proteins obtained from screening

Follow-up Validation Options for Yeast One-Hybrid Results

Service Content	Experimental Purpose	Details	Cycle (Working Days)
EMSA	Validate promoter and protein interaction	Probe synthesis and labeling	10
EMSA	Validate promoter and protein interaction	EMSA experiment	20
Dual-Luciferase Assay	Validate promoter and protein interaction	Vector construction	10-15
Dual-Luciferase Assay	Validate promoter and protein interaction	Dual-luciferase detection	25

Case Studies

Ready to Start Your Co-IP Project?

Our expert team is ready to assist you with your protein interaction research. Get in touch with us for a consultation or request a quote today.

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Q&A

Introduction to Yeast Hybrid-related vectors, strains, and their functions?

(1) pGBKT7 Vector: Used for constructing Nuclear Yeast Two-Hybrid Bait-pGBKT7 plasmids or as an empty control.
(2) pGADT7 Vector: Used for constructing Nuclear Yeast Two-Hybrid and One-Hybrid Prey-pGADT7 plasmids or as an empty control.
(3) pABAi Vector: Used for constructing One-Hybrid Bait-pABAi plasmids.
(4) pGBKT7-53 Plasmid & pGADT7-T Plasmid: Used as a positive control pair for Nuclear Yeast Two-Hybrid verification and screening.
(5) pGBKT7-lam Plasmid & pGADT7-T Plasmid: Used as a negative control pair for Nuclear Yeast Two-Hybrid verification and screening.
(6) pOST1-NubI Plasmid & pBT3-N-bait Plasmid: Used as a functional validation control pair for Membrane Yeast Two-Hybrid verification and screening.
(7) pBT3-N Vector: Used for constructing bait proteins with the N-terminus in the cytosol and C-terminus in the organelle lumen (or extracellular space).
(8) pBT3-SUC Vector: Used for constructing bait proteins with the N-terminus in the organelle lumen (or extracellular space) and containing an N-terminal signal peptide.
(9) pBT3-STE Vector: Used for constructing bait proteins with the C-terminus in the cytosol, N-terminus in the organelle lumen (or extracellular space), and without an N-terminal signal peptide.
(10) pBT3-N and pBT3-STE: Both can be used for bait proteins with both N- and C-termini located in the cytosol.
(11) pPR3-N Vector: Used for constructing prey proteins with the C-terminus in the cytosol and N-terminus extracellular or in the organelle lumen.
(12) pPR3-C Vector: Used for membrane library construction and prey proteins without transmembrane domains.
(13) Ptsu2-APP Plasmid & pNubG-Fe65 Plasmid: Used as a positive control pair for Membrane Yeast Two-Hybrid verification and screening.
(14) Ptsu2-APP Plasmid & pPR3-N Plasmid: Used as a negative control pair for Membrane Yeast Two-Hybrid verification and screening.
(15) Y187 Strain: A GAL4 system yeast strain developed by Clontech for One-Hybrid and Two-Hybrid experiments. MATα type. Can be directly transformed with plasmids or used for library screening via mating with MATa strains (Y2HGold, AH109, etc.). Genotype: trp1, leu2. Reporter genes: lacZ, MEL1.
(16) Y1HGold Strain: A GAL4-AbA yeast One-Hybrid system strain developed by Clontech. MATα type. Can be directly transformed with plasmids for library screening. Genotype: ura3, leu2. Reporter gene: AbAr.
(17) Y2HGold Strain: A GAL4 system yeast Two-Hybrid strain developed by Clontech. MATa type. Can be directly transformed with plasmids or used for protein interaction verification/library screening via mating with MATα strains like Y187. Transformation markers: trp1, leu2. Reporter genes: AbAr, HIS3, ADE2, MEL1.
(18) NMY51 Strain: A yeast Two-Hybrid strain developed by DUALsystems BioTech for detecting membrane protein interactions. Can be directly transformed with plasmids for protein interaction verification or library screening. Genotype: *trp1, leu2-3*. Reporter genes: HIS3, ADE2, lacZ.

How to choose between Yeast One-Hybrid and Yeast Two-Hybrid based on research needs?

Customer reviews

"This Y1H detection service is exceptional! High library coverage, low false positives, and accurate DNA-protein interaction identification. The process efficient, and the detailed report strongly supports our transcription factor research."

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