polymorphic Protocols

Category: Plant Biology Protocols: Plant Genetics Protocols

AFLP was designed as a highly sensitive method for DNA fingerprinting to be used in a variety of fields. We are using this technology to generate DNA based markers for cloning genes involved in phototropic responses in higher plants that have only been identified genetically by mutant phenotype. Protocol includes: Generate polymorphic recombinant F2 (or F3) population; Isolate genomic DNA; Restriction of DNA; Ligation of adapters; Pre-amplification of template DNA; AFLP-PCR; etc. - [Read AFLP For Positional Cloning]

Category: Genetics and Genomics: Cancer Genetics Protocols

Investigators can utilize X chromosome inactivation (methylation) to determine the clonality status of a tumor or premalignant lesion in females. The technique is based on a methylation-sensitive restriction enzyme and analysis of a polymorphic locus on the X chromosome. Clonal cell populations will show "loss" of the non-methylated allele after restriction digest. The assay can be performed on DNA recovered from microdissected samples. Both frozen tissue and fixed-embedded tissue can be used. - [Read Clonality - X Chromosome Inactivation Assay Protocol]

Category: Genetics and Genomics: Genotyping Protocols: PCR Genotyping

Plasmodium vivax is the second most prevalent malaria parasite affecting more than 75 million people each year, mostly in South America and Asia. In addition to major morbidity this parasite is associated with relapses and a reduction in birthweight. Practical PCR genotyping protocols based on polymorphic loci present in two P. vivax genetic markers. - [Read Practical PCR Genotyping Protocols for Plasmodium vivax using Pvcs and Pvmsp1]

Category: Fungi Protocols: Aspergillus Protocols

RAPD is a procedure for typing and fingerprinting isolates of a species. It can be used for epidemiological studies, such as investigations into hospital outbreaks and as a laboratory aid to keep track of cultures and to verify that mutants generated in the laboratory are genetically identical to the parental strain. In our hands, the use of one primer, R108, is sufficiently discriminatory to distinguish between the isolates of different strains. - [Read Random Amplification of Polymorphic DNA (RAPD) Typing and Fingerprinting Protocol]

Category: Yeast Protocols: Yeast Genetics Protocols: YACs Protocols

Protocol for the subcloning of Yeast artificial chromosome into phage lambda. To subclone the large insert fragment of human DNA contained within a YAC into a bacteriophage lambda vector. The subclones are 15 to 23 kb in size, and can be used to identify new polymorphic markers from a known region of the genome, to map a specific locus, and/or to screen other libraries. - [Read Subcloning of Yeast Artificial Chromosomes into Phage Lambda Protocol]