Purpose: The quality of DNA and reliability of molecular markers are crucial for the success of Polymerase Chain Reaction (PCR) based genetic sex determination. This study was aimed at investigating the optimum conditions for isolation of DNA from chicken blood and the reproducibility of dominant and co-dominant sex markers to be validated as a tool for successful sexing in avian research. Research Method: Efficacy of six different extraction procedures including manual and solution based commercial purification kit were evaluated with different combinations of initial blood, lysis buffer and protein denaturant in relation to the DNA yield and purity. Three primer sets namely CHD1, HUR 0423 and HUR 0424 were evaluated by PCR.
Findings: The study results showed that 10µl of initial blood volume yields a significantly high DNA yield with high purity. Dominant marker HUR0424 showed to be a reliable marker system for the genetic sexing of domestic chickens over co-dominant markers. Research Limitation: For the accuracy of the results, protocols had to be followed at the same time and using same sample to avoid any errors. Originality/ Value: PCR based sexing is considered, the most accurate and inexpensive method and hence validation of the method is important for success of future avian research.