mutation Protocols

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Assembling Aggregates between Diploid and Tetraploid Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4425

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes a method for producing diploid embryo-tetraploid embryo chimeras. It requires the timed combination of four-cell-stage tetraploid embryo production and the procedure for diploid embryo-diploid embryo aggregation. The resulting chimeras are useful for phenotypic analysis when an induced mutation has an extraembryonic phenotype. - [Read Assembling Aggregates between Diploid and Tetraploid Embryos Protocol]

Assembling Aggregates between Embryonic Stem (ES) Cells and Tetraploid Embryos Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/21/pdb.prot4426

Category: Mouse Protocols: Assembling Chimeras Mouse Protocols

Protocol describes a method for producing ES cell-tetraploid embryo chimeras. It requires the timed combination of four-cell-stage tetraploid embryo production and the procedure for ES cell-diploid embryo aggregation in which diploid embryos are replaced with tetraploid embryos. The resulting chimeras can be used to analyze the embryonic versus extraembryonic phenotype of a mutation. - [Read Assembling Aggregates between Embryonic Stem (ES) Cells and Tetraploid Embryos Protocol]

How to Determine Recessive-Lethal Mutation Rates - http://www.fgsc.net/neurosporaprotocols/How%20to%20determine%20recessive-leth%20final.pdf

Category: Fungi Protocols: Neurospora Protocols

Information and tips on how to determine recessive-lethal mutation rates. - [Read How to Determine Recessive-Lethal Mutation Rates]

In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI - http://www.cshprotocols.org/cgi/content/extract/2006/2/pdb.prot3813

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for in vitro mutagenesis using double-stranded DNA templates. Two oligonucleotides are used to prime DNA synthesis catalyzed by a high-fidelity thermostable polymerase on a denatured plasmid template. The two oligonucleotides both contain the desired mutation and occupy the same starting and ending positions on opposite strands of the plasmid DNA. - [Read In Vitro Mutagenesis Using Double-stranded DNA Templates: Selection of Mutants with DpnI]

Isolation of Arabidopsis cDNAs by Complementation in Yeast - http://www.arabidopsis.org/cshl-course/12-yeast.html

Category: Plant Biology Protocols: Arabidopsis Nucleic Acid Protocols

Includes: Isolation of Arabidopsis LEU2 cDNAs by complementation of the yeast leu2 mutation; Recovery of plasmid DNA from yeast cells; Analysis of Arabidopsis cDNAs that complement a yeast leu2 mutation; Preparation of library DNA from the Lacroute cDNA library; Preparation of Yeast Media. - [Read Isolation of Arabidopsis cDNAs by Complementation in Yeast]

Microinjection of Mouse ES Cells into Blastocysts - http://www.bcm.edu/dtmc/?pmid=7791

Category: Microinjection Protocols: Microinjection of DNA Protocols

The core defines one injection as a total of forty blastocysts injected on two consecutive days using one or two clones for the same mutation. Please, refer to the Pricing page for information on the cost of one injection. - [Read Microinjection of Mouse ES Cells into Blastocysts]

Mutation Detection By Single-Strand Conformational Polymorphism (SSCP) Protocol - http://www.ohsu.edu/nsi/faculty/reddyh/lab/protsscp.html

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

Protocol for mutation detection by single-strand conformational polymorphism (SSCP). Includes: PCR Protocol; SSCP Gels; - [Read Mutation Detection By Single-Strand Conformational Polymorphism (SSCP) Protocol]

Mutation Detection By Single-Strand Conformational Polymorphism (SSCP) Protocol - http://www.ohsu.edu/nsi/faculty/reddyh/lab/protsscp.html

Category: Genetics and Genomics: Cancer Genetics Protocols: Single Strand Conformational Polymorphism(SSCP) Protocols

Protocol for mutation detection by single-strand conformational polymorphism (SSCP). Includes: PCR Protocol; SSCP Gels. - [Read Mutation Detection By Single-Strand Conformational Polymorphism (SSCP) Protocol]

Mutation Detection by Solid-Phase Chemical Cleavage of Mismatches Using Radioactive Probes Protocol - http://www.cshprotocols.org/cgi/content/extract/2006/14/pdb.prot4120

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

This protocol is the first of two describing solid-phase chemical cleavage of mismatch (SpCCM), a method for the detection of mutations in genomic DNA. DNA fragments up to 2 kb in length can be analyzed to determine the position (within 10-15 bp) and identity of the mismatched nucleotide. - [Read Mutation Detection by Solid-Phase Chemical Cleavage of Mismatches Using Radioactive Probes Protocol]

Screening DNA Pools for T-DNA Insertions in Arabidopsis Genes Protocol - http://www.cshprotocols.org/cgi/content/abstract/2006/28/pdb.prot4622

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]

SNP Restriction Digestion Comparison - http://insilico.ehu.es/restriction/two_seq/

Category: DNA Protocols: SNP Protocols

Compare restriction patterns of two sequences for SNP/Mutation detection. Online program that compares restriction enzyme digest of DNA and finds the differences between two sequences. - [Read SNP Restriction Digestion Comparison]

Transposon Mediated Mutagenesis Protocol - http://www.genome.wisc.edu/resources/protocols/tnmutationprotocol.htm

Category: Cloning Protocols: Mutagenesis Protocols

Protocol for transposon mediated mutagenesis. Includes: Amplify ORF from MG1655; Transposition reaction; Transformation; Picking; Verify mutation by Culture PCR; Confirm mutations by sequencing; Curing the temperature sensitive pKD46 plasmid. - [Read Transposon Mediated Mutagenesis Protocol]

Transposon-Mediated Mutagenesis Protocol - http://www.genome.wisc.edu/resources/protocols/tnmutationprotocol.htm

Category: Bacteria Genetics Protocols

Protocol for transposon-mediated mutagenesis. Includes: Amplify ORF from MG1655; Transposition reaction; Transformation; Picking; Verify mutation by Culture PCR; Confirm mutations by sequencing; Curing the temperature sensitive pKD46 plasmid. - [Read Transposon-Mediated Mutagenesis Protocol]

Articles (1-2 of 2)

Electrotransformation of BMH 81-17mut S for Isolating Site-Directed dsDNA Mutants

This protocol describes the electroporation of the BMH 81-17 mut S strain that is recommended for tranformation of the site directed mutagenesis of dsDNA (See Protocol on Site-Directed Mutagenesis on Double Stranded DNA). BMH 81-17 mut S are a mismatch repair defective (mut S) Escherichia coli strain. The probability that the two mutations will cosegregate during the first round of DNA replication is increased in this strain.

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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