Genetic relationship between generations of chickens using RAPD-PCR

Abstract

     This search includes (180 male and 180 female) from the three generations (G) grandparent, (P) parent, and (B) broiler generations of commercial chicken (Ross). Blood samples were collected in 3 ml tubes with EDTA, genomic DNA extracted by Promega's Wizard Genomic DNA Purification Kit. The Nano Drop® spectrophotometer has been used to analyze DNA purity and concentration and it’s varied from (1.7 to 1.9). RAPD-PCR was performed using 18 primers from the GenScript USA Series (Table 1). The reaction mixture are 25 μl made up the PCR reaction. An initial phase of DNA denaturation at 94°C for 5 minutes was followed by 40 cycles of DNA denaturation at 94°C for 1 minute, annealing as described with each primer, extension at 72°C for 1 minute, and final extension at 72°C for 5 minutes. Electrophoresis on 2% agarose gels in 1x TBE buffer (Promega, USA). A UV transi was used to see the amplified pattern. The OPA-07 Primer produced the most bands (73) bands among all generations used. All of the Primers combined to generate a total of (442) bands, and there were 48 bands that were polymorphic. The primer OPQ-12 had the highest percentage of polymorphisms, (23.53), Primer OPA-07 has the widest molecular weight range (100-1500 bp). The highest genetic similarity is observed 0.813 between (P♀) and (B♂).The dendrogram consists of two clusters, one of which includes (B♀, P♀, B♂) and the other includes (G♂, G♀, P♂), where it is clear that the genetic distance is very high between (G♀ and P♀), while it is less between (B♂ and P♀) and also between (G♂ and G♀) while the distance is intermediate between (B♂ and B♀) and also between (G♂ and P♂). The goal of the current study was to use RAPD-PCR to determine the genetic distance and genetic relationship between generations of chickens.