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A Targeted Screen to Detect Recessive Mutations that have Quantitative Effects Protocol - http://www.nervenet.org/papers/Consomic.html

Category: Genetics and Genomics: Genotyping Protocols: Mutation Detection Protocols

Describes an experimental cross in mice that can be used to define and map induced germ-line mutations that map to a single chromosome. The cross is a modification and extension of a conventional three-generation recessive mutagenesis screen. Includes: The Mutagenesis Breeding Plan; Consomic Strains; Generating Mutations; Generating and Genotyping G2 Females; Genotyping G3 Progeny; Phenotyping G4 Progeny; etc.. - [Read A Targeted Screen to Detect Recessive Mutations that have Quantitative Effects Protocol]

Bacterial Cell Maintenance Protocol - http://www.genome.ou.edu/protocol_book/protocol_partI.html#I.G

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

4 strains of E. coli are used in these studies: JM101 for M13 infection and isolation, XL1BMRF'for M13 or pUC-based DNA transformation, and ED8767 for cosmid DNA transformation. To maintain their respective F' episomes necessary for M13 viral infection, JM101 is streaked onto a M9 minimal media plate and XL1BMRF' is streaked onto an LB plate containing tetracycline. ED8767 is streaked onto an LB plate. These plates are incubated at 37degC overnight. For each strain, 3 ml. of appropriate liquid.. - [Read Bacterial Cell Maintenance Protocol]

Curing Strains of Endogenous 2-Micron Plasmid Protocol - http://ygac.med.yale.edu/mtn/reagent/avail_reagents/curing2micron_info.stm

Category: Yeast Protocols: Yeast Genetics Protocols

Protocol for curing strains of endogenous 2-micron plasmid. - [Read Curing Strains of Endogenous 2-Micron Plasmid Protocol]

Delivery of dsRNA into Plants by VIGS Methodology - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4327

Category: Plant Biology Protocols: Plant Genetics Protocols

Virus-induced gene silencing (VIGS) uses a virus to deliver a sequence from a gene of interest into a host plant. The virus carrying the fragment of the gene of interest must be capable of replication if dsRNA is to be produced. One or two leaves are inoculated with Agrobacterium strains carrying the VIGS vector possessing the gene fragment. The virus then replicates and spreads throughout the plant, mediating silencing. - [Read Delivery of dsRNA into Plants by VIGS Methodology]

DNA Preparation Bacterial DNA from Mucoid Strains Protocol - http://rothlab.ucdavis.edu/protocols/D03.html

Category: Nucleic Acid Purification Protocols: DNA Isolation Protocols

Protocol for DNA preparation bacterial DNA from mucoid strains. - [Read DNA Preparation Bacterial DNA from Mucoid Strains Protocol]

Ethyl Methane Sulfonate (EMS) Mutagenesis Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/1/pdb.prot4180

Category: Yeast Protocols: Yeast Genetics Protocols

Protocol describes mutagenesis of yeast with ethyl methane sulfonate (EMS). It causes approximately 40-70% cell death in most haploid laboratory strains, a level of cell killing that is commonly used in mutant hunts with haploid strains. - [Read Ethyl Methane Sulfonate (EMS) Mutagenesis 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]

Genotyping Protocol for TgN(HLA-A2.1)1Enge - http://jaxmice.jax.org/pub-cgi/protocols/protocols.sh?objtype=protocol&protocol_id=442

Category: Molecular Model Organisms: Mouse Animal Protocols: Mouse Genotyping Protocols

24-mer A=4, C=10, G=2, T=8, HLA-A2.1 ( 003475) primer designed in Mouse Genotyping lab. ... This genotyping protocol is used for the following strains: ... - [Read Genotyping Protocol for TgN(HLA-A2.1)1Enge]

Genotyping Protocol: T1320 mouse - http://www.mmrrc.org/strains/425/ctr_protocol.pdf

Category: Molecular Model Organisms: Mouse Animal Protocols: Mouse Genotyping Protocols

Genotyping Protocol: MMRRC 425. Assay Type: PCR - can distinguish heterozygous animals from homozygous animals. DNA Extraction: DNA from tail snips was ... - [Read Genotyping Protocol: T1320 mouse]

Handling and Using Insertion Libraries - http://ygac.med.yale.edu/mtn/protocol.stm

Category: Yeast Protocols

Handling and using insertion libraries. Includes: Curing strains of endogenous 2-micron; mTn-lacZ/LEU2 insertion library; mTn-3xHA/lacZ insertion library; mTn-3xHA/GFP insertion library. - [Read Handling and Using Insertion Libraries]

How to Choose Wild Type Laboratory Strains of N. crassa for Reference and for Special Uses - http://www.fgsc.net/neurosporaprotocols/How%20to%20choose%20wild%20types%20for%20reference%20and%20special%20uses.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to choose wild type laboratory strains of N. crassa for reference and for special uses. - [Read How to Choose Wild Type Laboratory Strains of N. crassa for Reference and for Special Uses]

How to Obtain and Recognize Partial-Diploid Strains that are Duplicated for Known Chromosome Segment - http://www.fgsc.net/neurosporaprotocols/How%20to%20obtain%20and%20recognize%20par.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to obtain and recognize partial-diploid strains that are duplicated for known chromosome segments. - [Read How to Obtain and Recognize Partial-Diploid Strains that are Duplicated for Known Chromosome Segment]

How to Procure Neurospora Strains for Research or Teaching - http://www.fgsc.net/neurosporaprotocols/How%20to%20procure%20Neurospora%20strains%20for%20research%20and%20teaching.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to procure Neurospora strains for research or teaching. - [Read How to Procure Neurospora Strains for Research or Teaching]

How to Test for Complementation Between Mutant Strains - http://www.fgsc.net/neurosporaprotocols/How%20to%20test%20for%20complementation%20.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to test for complementation between mutant strains. - [Read How to Test for Complementation Between Mutant Strains]

How to Use Helper Strains for Maintaining and Crossing Handicapped Recessive Mutants - http://www.fgsc.net/neurosporaprotocols/How%20to%20use%20helper%20strains%20for%20maintaining%20and%20crossing%20handicapped%20recessive%20mutants,%20for%20forcing%20and%20resolving%20heterokaryons,%20and%20for%20determining%20heterokaryon%20compatibility

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to use helper strains for maintaining and crossing handicapped recessive mutants, for forcing and resolving heterokaryons, and for determining heterokaryon compatibility. - [Read How to Use Helper Strains for Maintaining and Crossing Handicapped Recessive Mutants]

How to Use Multiply Marked Multicent Strains for Mapping Genes and Translocation Breakpoints - http://www.fgsc.net/neurosporaprotocols/How%20to%20use%20multiply%20marked%20multicent%20strains%20for%20linkage%20group%20assignment%20of%20genes%20and%20rearrangement%20breakpoints.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to use multiply marked multicent strains for mapping genes and translocation breakpoints. - [Read How to Use Multiply Marked Multicent Strains for Mapping Genes and Translocation Breakpoints]

How to Work with Slime and Other Cell Wall-Deficient Strains - http://www.fgsc.net/neurosporaprotocols/How%20to%20work%20with%20slime%20final.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to work with slime and other cell wall-deficient strains. - [Read How to Work with Slime and Other Cell Wall-Deficient Strains]

Large-scale Preparation of Single-stranded and Double-stranded Bacteriophage M13 DNA Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3284

Category: Viral Manipulation Protocols: M13 Phage Manipulation Protocols

Protocol used chiefly to generate large stocks of double-stranded DNA of strains of M13 that are routinely used as cloning vectors. Large amounts of single-stranded DNA of an individual recombinant may occasionally be needed for specific purposes, e.g., to generate many preparations of a particular radiolabeled probe or to construct large numbers of site-directed mutants. - [Read Large-scale Preparation of Single-stranded and Double-stranded Bacteriophage M13 DNA Protocol]

Long Term Storage of Bacterial Strains Protocol - http://humgen.wustl.edu/hdk_lab_manual/plasmid/plsmid02.html

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

Bacterial strains may be stored indefinitely at low temperatures (- 20 degrees C and -80 degrees C) in 15 to 40% glycerol. - [Read Long Term Storage of Bacterial Strains Protocol]

Long Term Storage of Yeast Stocks Protocol - http://humgen.wustl.edu/hdk_lab_manual/yeast/yeast2.html

Category: Yeast Cell Culture Protocols: Yeast Media, Solutions and Stocks Protocols

Yeast strains may be stored indefinitely at low temperatures (-80 degrees C). Two archiving methods are presented below. In Method A , the cells are grown on a plate, while in Method B the cells are grown in liquid culture. - [Read Long Term Storage of Yeast Stocks Protocol]

Maintenance of Probes in Bacteria Including Escherichia coli Protocol - http://wheat.pw.usda.gov/~lazo/methods/lazo/bactstor.html

Category: Bacterial Cell Culture Protocols: Bacterial Stocks Protocols

Plasmid (pUC series) containing genomic DNA fragments are maintained in E. coli strain DH5aTM. The E. coli cultures are routinely cultured at 37 C on Luria-Bertani (LB) agar on or in LB broth containing Ampicillin (30 µg/ml) or Carbenicillin (50 µg/ml broth, 100 µg/ml agar). E. coli strains are usually preserved in stab agar or glycerol for mid-term storage and lyophilized for long-term storage. - [Read Maintenance of Probes in Bacteria Including Escherichia coli Protocol]

Methods for Maintaining Insect Cell Cultures - http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=355909

Category: Cell Culture Protocols: Insect Cell Culture Protocols

Insect cell cultures are now commonly used in insect physiology, developmental biology, pathology, and molecular biology. As the field has advanced from methods development to a standard procedure, so has the diversity of scientists using the technique. This paper describes methods that are effective for maintaining various insect cell lines. The procedures are differentiated between loosely or non-attached cell strains, attached cell strains, and strongly adherent cell strains. - [Read Methods for Maintaining Insect Cell Cultures]

Neurospora Methods Protocols - http://www.fgsc.net/methods/stanford.html

Category: Fungi Protocols: Neurospora Protocols

Protocols for Neurospora methods. Includes: Standard strains; Crosses; Minimal medium; Color coding of media; Agar substrate for manipulation and isolation; Stock solutions of supplements; Mating-type tests; Preservation of stocks by silica gel; Cleaning of glassware; Control of mites. - [Read Neurospora Methods Protocols]

PCR Genotyping of Mutant Strains - http://www.fhcrc.org/science/labs/soriano/protocols/pcrgen.html

Category: DNA Protocols: PCR Polymerase Chain Reaction Protocols

PCR Genotyping of Mutant Strains. Fred Hutchinson Cancer Research Center - [Read PCR Genotyping of Mutant Strains]

PCR Genotyping of Mutant Strains - http://www.fhcrc.org/science/labs/soriano/protocols/pcrgen.html

Category: PCR Protocols

PCR Genotyping of Mutant Strains. Fred Hutchinson Cancer Research Center - [Read PCR Genotyping of Mutant Strains]

Articles (1-2 of 2)

Constructing Random-Peptide Libraries for Phage Display

The protocol describes the construction of a large (10^8 to 10^10) primary random peptide phage library in fUSE5 or f88-4 strains.

Vector Design Protocol for Transgenic Mice Generation

The protocol gives general considerations for the design of targeting vectors for transgenic mice. The protocol shares tips in the design of knock-out and knock-in vectors and some of their strategies for producing homologously recombined embryonic stem cells.

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