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2 Hybrid System TRAFO Protocol - http://www.umanitoba.ca/faculties/medicine/biochem/gietz/2HS.html

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

Yeast Two-Hybrid System. TRAFO Solutions. Gietz Lab - [Read 2 Hybrid System TRAFO Protocol]

3'-End cDNA Amplification Using Classic RACE Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4130

Category: PCR Protocols: cDNA Synthesis Protocols

To generate "3'-end" partial cDNA clones, mRNA is reverse-transcribed using a "hybrid" primer (Qtotal, QT) that consists of two mixed bases (GATC/GAC followed by [T]17) and a unique 35-base oligonucleotide sequence (QI-QO). Amplification is then performed using a primer containing part of this sequence (Qouter, Qo) (which now binds to each cDNA at its 3'-end) and a primer derived from the gene of interest, GSP1 (gene-specific primer 1). - [Read 3'-End cDNA Amplification Using Classic RACE Protocol]

Colony Hybridization Protocol - http://cmgm.stanford.edu/~vesicle/protocols/hybrid.html

Category: RNA Protocols: cDNA Libraries Library

Methods and procedure for Colony Hybridization. Vesicle Trafficking Home. - [Read Colony Hybridization Protocol]

Directed Yeast Two Hybrid Screening for MEKK1 Interaction Partners Protocol - http://www.natureprotocols.com/2007/01/22/directed_yeast_two_hybrid_scre.php

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

Protocol utilizes the Y190 yeast strain, that characteristically turns from white to red during growth due to ademutation. Baits were subcloned into the pAS1cyh2 vector, and Preys subcloned into pACT2. - [Read Directed Yeast Two Hybrid Screening for MEKK1 Interaction Partners Protocol]

EXAMINING THE FUNCTION OF PROTEINS AND PROTEIN NETWORKS WITH THE YEAST TWO-HYBRID SYSTEM - http://proteome.wayne.edu/Update.html

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

Interactor Hunt Protocol. Testing whether a bait activates transcription. Screening a library for interactors. Finley Lab. 1997. - [Read EXAMINING THE FUNCTION OF PROTEINS AND PROTEIN NETWORKS WITH THE YEAST TWO-HYBRID SYSTEM]

Gateway-Compatible Yeast One-Hybrid Screens Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/28/pdb.prot4590

Category: Yeast Protocols: Yeast Nucleic Acid Protocols

Protocol for gateway-compatible yeast one-hybrid screens. - [Read Gateway-Compatible Yeast One-Hybrid Screens Protocol]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4562

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This procedure capitalizes on the asexual/sexual reproductive cycle of yeast cells. Laboratory yeast strains can be maintained indefinitely by asexual reproduction as one of two haploid mating types, MATa or MAT{alpha}. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4563

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This protocol describes how to handle arrayed yeast using robotics. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array:]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: Amplification of ORFs - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4561

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This protocol describes the first step in constructing an array: amplification of the predicted ORFs that are to be included in the array. Gene-specific primers containing vector-specific flanking sequences that facilitate recombinational cloning are used to amplify each ORF. A secondary amplification can be used to extend the length of the homologous vector sequence flanking the ORF. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: Amplification of ORFs]

Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: S - http://www.cshprotocols.org/cgi/content/abstract/2006/23/pdb.prot4564

Category: Yeast Protocols: Yeast Protein Protocols: Yeast Protein Protein Interactions Protocols

This protocol describes three different storage methods for the array. Frozen stocks are the most permanent method and are an absolute requirement for maintaining a record of the array. - [Read Genome-Wide Analysis of Protein-Protein Interactions Using a Two-Hybrid Array: S]

Hybridization and Detection Using the HC ExpressArray Kit Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4122

Category: Microarray Protocols: Microarray Hybridization Protocols

Protocol describes the direct detection of RNA on DNA microarrays using Hybrid Capture (HC) technology and the HC ExpressArray Kit developed by Diagene. The kit uses a proprietary antibody that binds specifically to RNA:DNA hybrids and a second, fluorescently labeled, antibody that detects the primary antibody. Total RNA is applied directly to a glass-spotted DNA microarray, and stable RNA:DNA hybrids are visualized via a Cy3-labeled secondary antibody. - [Read Hybridization and Detection Using the HC ExpressArray Kit Protocol]

Isolation of Plant Transcription Factors Using a Modified Yeast One-Hybrid System - http://www.plantmethods.com/content/2/1/3

Category: Plant Biology Protocols: Plant Genetics Protocols

The preparation of expressional cDNA libraries for use in the yeast two-hybrid system is quick and efficient when using the dedicated Clontech™ product, the MATCHMAKER Library Construction and Screening Kit 3. This kit employs SMART technology for the amplification of full-length cDNAs, in combination with cloning using homologous recombination. - [Read Isolation of Plant Transcription Factors Using a Modified Yeast One-Hybrid System]

Two Hybrid System Transformation Protocol - http://www.umanitoba.ca/faculties/medicine/biochem/gietz/2HS.html

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

High Efficiency Transformation of a yeast strain requiring maintenance of a plasmid. - [Read Two Hybrid System Transformation Protocol]

Two-hybrid analysis of genetic regulatory networks - http://proteome.wayne.edu/YTHnetworks.html

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

Finley and Brent. Excellent Two-Hybrid Resource Article and Protocols. Very Detailed. - [Read Two-hybrid analysis of genetic regulatory networks]

Two-hybrid Systems Stage 1: Characterization of a Bait-LexA Fusion Protein Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3895

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

Protocol describes how to generate a plasmid construct (pBAIT) that expresses a target protein fused to the bacterial LexA protein. PBAIT is cotransformed into yeast with a lexAop-lacZ reporter plasmid carrying the bacterial lacZ gene under the control of the lexA operator. The recipient yeast strain contains a chromosomally integrated leu2 reporter gene, also under the control of the lexA operator. - [Read Two-hybrid Systems Stage 1: Characterization of a Bait-LexA Fusion Protein Protocol]

Two-hybrid Systems Stage 2: Selecting an Interactor Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3888

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

In this stage of the protocol, a mammalian cDNA library constructed in a plasmid such as pJG4-5 is transformed into yeast strains containing pBAIT and the lexAop-lacZ reporter plasmid. PJG4-5 expresses the cloned cDNAs from a cassette containing a transcriptional activation domain and other moieties under the control of the yeast GAL1 promoter. - [Read Two-hybrid Systems Stage 2: Selecting an Interactor Protocol]

Yeast Two Hybrid Assay: A Fishing Tale - http://www.bioteach.ubc.ca/Journal/V01I01/8188yeast.pdf

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

Theory, Galactose metabolism, The Pros and Cons of the Two hybrid Assay. Troubleshooting and Tips for Hybrid assay. Sobhanifar BioTeach Journal 2003. - [Read Yeast Two Hybrid Assay: A Fishing Tale]

Yeast Two-Hybrid Screen with Library and Bait - http://faculty.virginia.edu/mirmira/resources_files/Protocols/YEAST%20TWO.htm

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

yeast transformed with bait vector. yeast culture. selection plates. Mirmira, Univ. Virginia. - [Read Yeast Two-Hybrid Screen with Library and Bait]

Yeast Two-hybrid Systems Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3889

Category: Yeast Protocols: Yeast Two-Hybrid Systems Protocols

Protocol for Yeast two-hybrid systems. - [Read Yeast Two-hybrid Systems Protocol]

Yeast Two-Hybrid: State of the Art - http://www.biologicalprocedures.com/bpo/arts/1/16/m16.htm

Category: Molecular Model Organisms: Yeast Two-Hybrid Screen Protocol

Yeast Two-Hybrid: State of the Art. Paper on Protein-protein interactions and the two-hybrid protocol. Wim Van Criekinge et al., BPO. - [Read Yeast Two-Hybrid: State of the Art]

Articles (1-3 of 3)

Genomic DNA Labeling of Microarrays Protocol

DNA microarrays are an ordered arrangement of DNA molecules complementary to genes of interest that are "spotted" by robotic equipment onto a glass slide substrate. The expression of genes in cells can be monitored with microarrays by preparing cDNA from the mRNA of cells of interest and measuring the hybridization to the microarray. This protocol describes the labeling of genomic DNA for use as a probe for hybridization to the cDNA spotted on the array.

Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA

This Microarray Protocol Preparation of Fluorescent DNA Probes from Human mRNA protocol describes the production of probes labeled with the fluorescent dyes, Cy3 and Cy5, following the synthesis of cDNA from human mRNA and the hybridization of the probes to DNA microarrays.