A quantitative assay for crossover and noncrossover molecular events at individual recombination hotspots in both male and female gametes.
Document Type
Article
Publication Date
2008
Keywords
Chromosomes-Mammalian, Cloning-Molecular, Crossing-Over-Genetic, Female, Male, Meiosis, Mice, Oocytes, Polymorphism-Single-Nucleotide, Spermatozoa
First Page
204
Last Page
209
JAX Source
Genomics 2008 Oct; 92(4):204-9.
Abstract
Meiotic recombination is a fundamental process in all eukaryotes. Among organisms in which recombination initiates prior to synapsis, recombination preferentially occurs in short 1-to 2-kb regions, known as recombination hotspots. Among mammals, genotyping sperm DNA has provided a means of monitoring recombination events at specific hotspots in male meiosis. To complement these current techniques, we developed an assay for amplifying all copies of a hotspot from the DNA of male and female germ cells, cloning the products into Escherichia coli, and SNP genotyping the resulting colonies using fluorescence technology. This approach examines the molecular details of crossover and noncrossover events of individual meioses directly at active hotspots while retaining the simplicity of using pooled DNA. Using this technique, we analyzed recombination events at the Hlx1 hotspot located on mouse chromosome 1, finding that the results agree well with a prior genetic characterization of 3026 male and 3002 female meioses.
Recommended Citation
Ng SH,
Parvanov E,
Petkov PM,
Paigen K.
A quantitative assay for crossover and noncrossover molecular events at individual recombination hotspots in both male and female gametes. Genomics 2008 Oct; 92(4):204-9.