The present study was carried out for investigation of polymorphism of FecB and POU1F1 gene in Assam Hill goat. Blood samples pertaining to 80 randomly selected Assam Hill goats having kidding history of single as well as multiple births maintained at three field units viz., Batabari, Nahira and Tetelia under “AICRP on Goat Improvement” were utilized. DNA was extracted using modified phenol chloroform extraction procedure. The quantity and quality of extracted DNA were assessed using spectrophotometry and agarose gel electrophoresis. A 190 bp fragment of FecB gene and a 450 bp fragment of POU1F1 gene were amplified using Polymerase Chain Reaction (PCR). RFLP analysis of FecB using AvaII enzyme revealed undigested 190 bp product for all the samples. Digestion of POU1F1 with DdeI produced three fragments of 102 bp, 118 bp and 200 bp in agarose gel electrophoresis for all the samples revealing monomorphism. The POU1F1 PCR-RFLP products were also visualized by loading on 12% PAGE in TBE buffer which resulted in similar banding patterns. RFLP analysis of POU1F1 gene was also performed by using PstI, resulting in undigested product of 450 bp. In sequence analysis, no restriction site was found for AvaII in FecB gene and four restriction sites were found for DdeI in POU1F1 gene. Sequence analysis of the samples revealed 99-100% homology for both the genes among all the samples irrespective of litter size. | Investigation of FecB and POU1F1 gene polymorphism in Assam hill goat