clones Protocols

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: PCR Polymerase Chain Reaction Protocols

Identification of targeted ES cell clones by PCR. Fred Hutchinson Cancer Research Center. - [Read PCR Screens of ES Cell Clones]

Category: PCR Protocols

Identification of targeted ES cell clones by PCR. Fred Hutchinson Cancer Research Center. - [Read PCR Screens of ES Cell Clones]

Category: Viral Culture and Isolation Protocols: Viral Culture Protocols

This rapid method is used to screen bacteriophage {lambda}gt11 clones for the production of immunodetectable fusion proteins. After optimizng the conditions of infection and induction, the method can be used to produce preparative amounts of a fusion protein. - [Read Preparation of Lysates Containing Fusion Proteins Encoded by Bacteriophage {lambda}: Lytic Infection]

Category: Immunology: Mononuclear Cell Isolation Protocols: T-Cell Isolation Protocols

Protocols utilize T hybridomas to detect expression of peptide-MHC complexes, since these cells provide the most convenient, consistent, and flexible T cell readout systems for these purposes. If desired, antigen-specific T cell clones can be used in lieu of T hybridoma cells, but T cell clones often give poorer responses than T hybridomas to fixed APCs due to fixation-induced loss of costimulator function. - [Read Presenting Exogenous Antigen to T Cells Protocol]

Category: Immunology: T Cell Protocols

Provides methods for the derivation of specific types of T cell clones: preparation and maintenance of alloreactive murine helper T (TH) lymphocyte and cytotoxic T lymphocyte (CTL) clones using the limiting dilution technique and derivation of TH clones reactive with soluble protein antigens including a method for the selection of either TH1 or TH2 lymphocyte subsets. - [Read Production of T Cell Clones 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: PCR Protocols: cDNA Synthesis Protocols

This method is used to extend partial cDNA clones by amplifying the 5' sequences of the corresponding mRNAs.  The technique requires knowledge of a small region of sequence within the partial cDNA clone.  During PCR, the thermostable DNA polymerase is directed to the appropriate target RNA by a single primer derived from the region of known sequence. - [Read Rapid Amplification of 5' cDNA Ends 5'-RACE Protocol]

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 sizing of YAC clones. To determine the size of a yeast artificial chromosome within the background of the normal yeast chromosomal complement. - [Read Sizing of YAC Clones 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: Pharmacology and Toxicology Protocols: Receptor Binding Protocols

Stimulation of MHC class I binding receptors on decidual NK clones. - [Read Stimulation MHC Class I Binding Receptors]

Category: Immunology: T Cell Protocols

Protocol for stimulation of MHC class I binding receptors on decidual NK clones. - [Read Stimulation of MHC Class I Binding Receptors on Decidual NK Clones Protocol]

Category: Viral Manipulation Protocols: Phage Protocols

Protocol for transfer of P1 clones by electroporation. - [Read Transferring Bacteriophage P1 Clones between E. coli Hosts Protocol]

Category: Viral Manipulation Protocols: Phage Protocols

Protocol describes methods for recovery and purification of recombinant clones of bacteriophage P1 or PAC DNAs from bacteria. Because of their large size, these DNAs are sensitive to shearing forces and must be handled carefully. This protocol generally yields P1 DNA that works well as a substrate or template in enzymatic reactions. - [Read Working with Bacteriophage P1 and Its Cloning Systems Protocol]