Development of the arbitrarily primed-representational difference analysis method and chromosomal mapping of isolated high throughput rat genetic markers (arbitrarily primed–PCRypolymorphic markersyrat genome)
نویسندگان
چکیده
Linkage mapping of quantitative trait loci requires analysis of a large number of animals. Although genetic markers isolated by representational difference analysis (RDA) and its modifications meet the needs, the number of these markers has been limited. In the present study, we established the arbitrarily primed (AP)–RDA method to isolate virtually an unlimited number of the high throughput genetic markers. A representation of the genome, an APamplicon, was prepared by AP-PCR with a single primer or with a combination of primers using genomic DNA of the ACIyN (ACI) or BUFyNac (BUF) rat as a template. By subtracting the AP-amplicon of ACI from that of BUF, a total of 40 polymorphic and independent markers were isolated in seven series of AP-RDA using a single primer. Two series of AP-RDA with primer combination yielded seven additional independent markers. All of the markers gave clear positivey negative signals by hybridization of a filter where APamplicons from F2 rats of ACI and BUF were dot-blotted at a high density without any concentration or purification. All of the 47 independent markers were mapped to unique chromosomal positions by linkage analysis, even though some arbitrary primers had very similar sequences. The markers were also informative between other strains of rats. Simultaneous hybridization of multiple filters made it possible to genotype a large number of rats simultaneously for multiple genetic loci. The AP-RDA method promises isolation of a large number of high throughput genetic markers in any species and is expected to facilitate linkage mapping of subtle quantitative
منابع مشابه
I-37: Genome Instability and DNA Damage in Male Somatic and Germ Cells Expressed as Chromosomal Microdeletion and Aneuploidy Is A Major Cause of Male Infertility
Background: Sperm chromatin insufficiencies leading to low sperm count and quality, infertility and transmission of chromosomal microdeletion and aneuploidies to next generations can be due to exposure to environmental pollutions, chemicals and natural or manmade ionizing radiation. In this project which has continued for more than 10 years and is unique in many technical aspects in Iran and in...
متن کاملAnalysis of Genomic Instability and Tumor-Specific Genetic Alterations by Arbitrarily Primed PCR
متن کامل
Application of DNA Molecular Markers in Plant Breeding (Review article)
Plant Breeding has utilized a wide range of techniques and methods to improve the quality and quantity of plants. The molecular markers are the tools that have provided a new perspective for plant breeding advancements. This article has reviewed the various advantages and uses of molecular markers and the utilization of the high potential of natural polymorphisms within communities, combined wi...
متن کاملA rat genetic map constructed by representational difference analysis markers with suitability for large-scale typing.
Representational difference analysis (RDA) was applied to isolate chromosomal markers in the rat. Four series of RDA [restriction enzymes, BamHI and HindIII; subtraction of ACI/N (ACI) amplicon from BUF/Nac (BUF) amplicon and vice versa] yielded 131 polymorphic markers; 125 of these markers were mapped to all chromosomes except for chromosome X. This was done by using a mapping panel of 105 ACI...
متن کاملPotential Therapeutic Effect of TLR4-Primed Mesenchymal Stem Cells in Lessening Kidney Damages in Rat Model of Diabetic Nephropathy
Background and Aims: Substantial damage to the kidney tissue and diabetic nephropathy (DN) can be caused by chronic hyperglycemic conditions and exposure to a high level of blood glucose. In the current study, we explored the capability of adipose-derived mesenchymal stem cells (ADSCs) and Toll-like receptor-4-primed mesenchymal stem cells (TLR4-primed MSCs) on kidney regeneration, resolution ...
متن کامل