screening Protocols

Category: Plant Biology Protocols: Arabidopsis Genetics Protocols

A powerful way to identify a mutation in the gene of interest and to test mutant plants for phenotypes that are predicted to result from loss of function of that gene is by PCR screening. Pools of insertion lines are screened using one primer corresponding to the gene of interest and one primer corresponding to the end of the insertion element. The synthesis of a product indicates the presence of an insertion in the gene of interest. - [Read Screening DNA Pools for T-DNA Insertions in Arabidopsis Genes Protocol]

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

An expression library constructed in a bacteriophage {lambda} vector is plated on an appropriate E. coli strain in the absence of isopropylthio-ß-D-galactoside (IPTG). After 2-4 hours, the plates are moved to 37°C (to stabilize any fusion proteins that are temperature sensitive), and filters impregnated with IPTG are laid on top of the developing plaques. - [Read Screening Expression Libraries Constructed in Bacteriophage Lambda Vectors Protocol]

Category: DNA Protocols: cDNA Library Protocols

A cDNA library constructed in a plasmid expression vector of the pUC, pUR, or pEX series is plated on agar medium and then replicated onto filters, which are transferred to plates containing IPTG. After 2-4 hours of induction, the colonies are lysed with chloroform and then screened with appropriate antibodies. - [Read Screening Expression Libraries Constructed in Plasmid Vectors Protocol]

Category: Chromatography Protocols: Dye Ligand Affinity Chromatography Protocols

Protocol for screening immobilized dyes for their ability to bind a target protein. The selection of a dye adsorbent for the purification of the target protein is an empirical undertaking and is generally achieved by a trial-and-error approach. - [Read Screening Immobilized Dyes for their Ability to Bind a Target Protein Protocol]

Category: RNA Protocols: cDNA Libraries Library

General principles and procedures. Good tips. Atle M. Bones Lab. - [Read Screening of cDNA and genomic libraries.]

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Plaques formed by M13 bacteriophages or bacterial colonies transformed by plasmids carrying specific mutations can be detected by hybridization, using a radiolabeled oligonucleotide that forms a perfect duplex with the mutant sequence. Hybridization is carried out under conditions of low stringency that allow the radiolabeled oligonucleotide to anneal to both mutant and wild-type DNAs. - [Read Screening Recombinant Clones for Site-directed Mutagenesis by Hybridization to Radiolabeled Oligos]

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol for screening YAC libraries. - [Read Screening YAC Libraries Protocol]

Category: Antibody Protocols: Antibody Production Protocols

Protocol for steps in hybridoma production. Includes: SIMPLE METHOD: BY FROM GIBCO the HAT and HT concentrate; INSTRUMENTS FOR SPLENECTOMY OF ONE MICE; PREPARATION FOR CELL FUSION; CELL FUSION; SCREENING THE CLONES; RECLONING; ASCITIC FLUID PRODUCTION; PREPARATION OF PERITONEAL MACROPHAGES. - [Read Steps in Hybridoma Production Protocol]

Category: DNA Protocols: cDNA Library Protocols

Protocol describes the generation of cDNA probes from poly(A)+ mRNA using random oligonucleotide primers. Probes of this type are used for differential screening of cDNA libraries. - [Read Synthesis of cDNA Probes from mRNA Using Random Oligonucleotide Primers Protocol]

Category: RNA Protocols: cDNA Libraries Library

Munroe et al PNAS 1995 "..developed a PCR-based method for rapid and effective screening of arrayed cDNA libraries. - [Read Systematic screening of an arrayed cDNA library by PCR PDF]

Category: Transgenic Mouse Protocols

Clone the Transgene, transgene Screening, Transgene Expression, DNA microinjection, Identify Transgenic Founders, Transgenic Analysis. Transgenic Animal Web. - [Read Transgenic Outline: Mice and Rats]