screening Protocols

Category: Immunology: Immunoprecipitation Protocols

Protocol for immunoprecipitation of mRNA-protein complexes. In this protocol, an antibody targeting an RBP of interest is used to immunoprecipitate the RBP and any interacting molecules from a cell lysate. Reverse transcription followed by PCR is then used to identify individual mRNAs isolated with the RBP. This method focuses on examining an association between a specific RBP-mRNA complex, and it is best suited for a small scale screening of known or putative binding partners. - [Read Immunoprecipitation of mRNA-Protein Complexes Protocol]

Category: RNA Protocols: cDNA Libraries Library

Adsroption of Anti-E. coli Antibodies, cDNA Library Screening, Frank Lab. Oklahoma Medical Research Foundation - [Read Immunoscreening of Lambda-GT11 Bacteriophage Libraries with Antibody Probes]

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]

Category: Viral Manipulation Protocols: Phage Lamda Manipulation Protocols

Protocol for lambda gt22a screening. Includes: T-TYN Media + Mg+2; T-TYN Plates; T-TYN Top Agarose; Phage Dilution Buffer (PDB); 10X Kinase Buffer (-DTT); Calf Thymus DNA. - [Read Lambda gt22a Screening Protocol]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols: Liver Toxicity Assays Protocols

Leakage of lactate dehydrogenase and alanine aminotransferase from rat and mouse liver slices exposed to the test compound is used as a measure of hepatotoxicity. - [Read Liver Slice Hepatotoxicity Screening System Protocol]

Category: Microsphere Analysis Protocols

The role of microspheres in these screens is similar to their traditional role in immunoassays, namely as a solid phase to either enhance detection, separation, or both. The predominance of radioactive assays in high-throughput screening, along with the desire to find alternative means of detection, have led to research on substituting alternative fluorescent technologies. - [Read Microspheres for High-Throughput Screening Assays]

Category: Pharmacology and Toxicology Protocols: Cytotoxicity Protocols

Molecular and genetic toxicology studies on gene mutation in mammalian cells In vitro. Includes: G201 Mouse Lymphoma L5178Y/tk+/Cell Gene Mutation Assay; G201-R; G202 Mouse Lymphoma L5178Y/tk+/Cell Gene Mutation Screening Assay; G203 Chinese Hamster Ovary or V79 Cell Mutation Assay at the hprt Locus; G203R. - [Read Molecular and Genetic Toxicology Studies on Gene Mutation in Mammalian Cells In Vitro]

Category: ELISA Protocols

Protocol for monoclonal antibody screening by direct ELISA. - [Read Monoclonal Antibody Screening by Direct ELISA Protocol]

Category: DNA Protocols: SNP Protocols

MULTIPLEX SNP ANALYSIS: SCREENING FACTOR V R506Q (LEIDEN) MUTATIONS. Beckman Coulter. - [Read MULTIPLEX SNP ANALYSIS: SCREENING FACTOR V R506Q (LEIDEN) MUTATIONS]

Category: RNA Protocols: cDNA Libraries Library

Denaturation Solution, Neutralization Solution, Rinse Solution, Crosslink DNA to membranes using Stratalinker UV crosslinker. Donelson Lab, Univ. Iowa. - [Read Nitrocellulose Membrane Washing Procedure for cDNA Screening]

Category: Pharmacology and Toxicology Protocols: Enzyme Assays Protocols

Protocol for P450 screening system. - [Read P450 Screening Systems Protocol]

Category: RNA Protocols: cDNA Libraries Library

"useful screen phage library non-radioactive, and not need too many clones. Up to 5 clones can be obtained usually within one week." Methods.info - [Read PCR based Lambda λDNA library screening method]

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

PCR can be used to identify rare DNA sequences in DNA libraries by increasing the abundance of a particular sequence. This is accomplished by subdividing the original library into pools of decreased complexity and screening each pool or group of pools for a given DNA sequence. - [Read PCR-Based Screening of DNA Libraries Protocol]

Category: PCR Protocols

PCR can be used to identify rare DNA sequences in DNA libraries by increasing the abundance of a particular sequence.  This is accomplished by subdividing the original library into pools of decreased complexity and screening each pool or group of pools for a given DNA sequence.  A pool that contains the desired clone is subsequently subdivided into smaller pools, each of which is screened using the same PCR protocol that was used for the primary screen. - [Read PCR-Based Screening of DNA Libraries Protocol]

Category: Immunology: T Cell Protocols

T cell hybridomas can be obtained by fusing activated T cells with tumor cells. A heterogeneous population of hybridomas can be cloned by limiting dilution to obtain hybridomas that express specificity to one T cell receptor (TCR). Protocol describes cell fusion and selection of T cell hybridomas. A protocol is provided for screening of T cell hybridomas for expression of the CD3-TCR complex by flow cytometry analysis. - [Read Production of Mouse T Cell Hybridomas Protocol]

Category: Fungi Protocols: Neurospora Protocols

Protocol guide for the N. crassa yeast artificial chromosome library. Includes: Chromosome Walking; Hybridization screening of the YAC library; YAC restriction mapping and contig building; Preparation of chromosomal DNA plugs of YAC clones; Partial restriction enzyme digestion of YAC DNA plugs; Using CHEF gel analysis to resolve YAC clones; Southern Hybridization; Isolation of terminal restriction fragments from cloned DNA inserts in YAC clones; etc. - [Read Protocol Guide for the N. crassa Yeast Artificial Chromosome Library]

Category: RNA Protocols: cDNA Libraries Library

Hoskins et al. 2005 NAR. "... describe a PCR-based method for directed screening of plasmid cDNA libraries. " - [Read Rapid and efficient cDNA library screening by self-ligation of inverse PCR products (SLIP)]

Category: siRNA and RNAi Protocols

The protocol listed is Drosophila cells in 6 well plates and our pre-aliquoted 384 well plates. RNAi experiments may be done in other size plates, just scale up or down. Drosophila RNAi Screening Center - [Read RNA Interference Screening (RNAi) in Cultured Cells]

Category: RNA Protocols: cDNA Libraries Library

Procedures: Titer the library to determine the concentration of infectious phage, Plate and lift the phage particles, Hybridize the immobilized cDNA with labeled probe, Pick positive plaques. Karl Clark U. Minn. - [Read Screening a cDNA Library for use with HybriZAP zebrafish cDNA libraries]

Category: RNA Protocols: cDNA Libraries Library

Frank Lab. Oklahoma Medical Research Foundation. How to screen a Lambda Bacteriophage Genomic Library with a DNA Probe. - [Read Screening a Lambda Bacteriophage Genomic Library with a DNA Probe]

Category: DNA Protocols: Cosmid Library Protocols

Unamplified cosmid libraries are plated at high density onto 150-mm nitrocellulose or nylon filters and screened by hybridization - [Read Screening an Unamplified Cosmid Library by Hybridization: Plating the Library onto Filters Protocol]

Category: Bacterial Protocols

Bacterial colonies growing on agar plates are transferred en masse to nitrocellulose filters. The spatial arrangement of colonies on the plates is preserved on the filters. After transfer, the filters are processed for hybridization to an appropriate radiolabeled probe while the original (master) plate is incubated for a few hours to allow the bacterial colonies to regrow in their original positions. - [Read Screening Bacterial Colonies by Hybridization: Intermediate Numbers Protocol]

Category: Bacterial Protocols

This procedure is used to plate, replicate, and subsequently screen large numbers of bacterial colonies (up to 2 x 104 colonies per 150-mm plate or 104 colonies per 90-mm plate). - [Read Screening Bacterial Colonies by Hybridization: Large Numbers Protocol]

Category: Bacterial Protocols

Protocol used to screen a small number of bacterial colonies (<200) that are dispersed over several agar plates and are to be screened by hybridization to the same radiolabeled probe. The colonies are gridded onto a master plate and onto a nitrocellulose or nylon filter laid on the surface of a second agar plate. - [Read Screening Bacterial Colonies by Hybridization: Small Numbers Protocol]

Category: DNA Protocols: cDNA Library Protocols: cDNA Library Construction Protocols

Two methods of screening pools of cloned cDNAs are discussed: transfection into mammalian cells and injection into Xenopus oocytes. - [Read Screening cDNA Libraries Constructed in Eukaryotic Expression Vectors Protocol]