Services
Providing Advanced Biological Technical Services for Researchers in Fundamental Science
Yeast Library Construction Service (Gateway Technology)
Comprehensive Gateway-Based Yeast Library Solutions for High-Efficiency Screening
Overview
Our Yeast cDNA Library Construction Service using Gateway Technology offers a powerful and efficient platform for constructing high-quality yeast libraries compatible with a wide range of protein interaction screening applications. This advanced cloning method, originally developed by Invitrogen in 1999, leverages the site-specific recombination system of bacteriophage lambda, enabling seamless DNA fragment transfer between various vectors without the need for restriction enzymes or ligation.
We offer two types of Gateway-compatible yeast libraries: Nuclear System cDNA Libraries and Membrane System cDNA Libraries, each tailored for downstream yeast two-hybrid (Y2H) or membrane yeast two-hybrid (MYTH) screening workflows.
Key Advantages
· High-Quality RNA Extraction: Multiple validated protocols for various sample types ensure optimal RNA quality and library integrity.
· Strict Quality Control: Libraries undergo PCR validation and sequencing to confirm insert sizes and species accuracy.
· Customizable Library Vectors: We offer vector flexibility (e.g., pGADT7, pPR3-N) to support different screening needs.
· Post-Delivery Support: Professional guidance for downstream screening applications is provided.
· One-Year Free Warranty: Includes protection for shipping damage, improper storage, and accidental sample loss for peace of mind.
Service Workflow and Deliverables
Nuclear System Yeast cDNA Library (Gateway Technology)
Turnaround Time: 17 business days
Deliverables:
4 mL of primary and secondary library glycerol stocks (4 tubes each)
200 µg of primary library plasmid DNA (2 tubes)
400 µg of secondary library plasmid DNA (4 tubes)
Digital project report (Word format) with full data summary
Library quality guaranteed:
Library size >1×10⁷ CFU
Average insert size: 0.8–1.5 kb (species-dependent)
Empty vector rate <5%
Library Construction Steps:
1. Total RNA extraction
2. mRNA purification
3. First-strand and second-strand cDNA synthesis
4. Adapter ligation with attB1 sequences
5. cDNA size fractionation
6. BP recombination with pDONR222
7. DH10B E. coli transformation
8. Library validation and plasmid extraction
9. LR recombination with pGADT7
10. Final transformation and validation
11. Data analysis and reporting
Membrane System Yeast cDNA Library (Gateway Technology)
Turnaround Time: 17 business days
Deliverables: Same as Nuclear Library, except final vector is pPR3-N for membrane protein interaction studies.
Library Construction Steps: Same as above, with LR recombination into pPR3-N for compatibility with membrane Y2H systems.
Sample Collection Guidelines
To ensure the integrity of your cDNA library, we recommend following these best practices:
Sample Preservation
Process samples quickly to minimize RNA degradation.
For fresh tissues, isolate samples on ice and store immediately in liquid nitrogen, dry ice, or at -80°C.
Sample Shipping
Use aluminum foil or centrifuge tubes for packaging.
Label samples clearly with waterproof markers.
Ship with dry ice (5–30 kg depending on transit time) in a sealed foam box.
Sample Requirements
Sample Type | Minimum Amount |
|---|---|
RNA | >400 µg total RNA |
Plant Tissues | >10 g |
Fruits | >50 g |
Animal Tissues/Insects | >10 g |
Fungi | >10 g |
Cells / PBMC | >1×10⁸ cfu |
Case Results
Application Areas
· Protein-protein interaction mapping
· Functional genomics
· Novel gene discovery
· Membrane protein screening (MYTH)
· Signal transduction analysis
Disclaimer: All products and services are intended for research use only. They are not for diagnostic or therapeutic purposes.
Services Workflow
Online Consultation
01
Solution Matching
02
Service Contract
03
One-Stop-Services
04
Project Report
05
Related Products
Product Inquiry
Contact Us
Tel: +86 025-85205672
Email: info@pronetbio.com
Address: Building 3C, Nanjing Xianlin Zhigu,
Qixia District, Nanjing, Jiangsu, China, 210033
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