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Not AvailableToll-like receptors are one of the pattern recognition receptors important in the innate immunity of vertebrates. TLR4 is well known for recognition of LPS of gram negative bacteria leading to production of inflammatory cytokines. MyD88 is one of the adaptor molecules facilitating downstream signalling in TLR pathway. The aim of this study was to design PCR primers and standardize PCR conditions for amplification of Toll-like receptor 4 and MyD88 in Pangasianodon hypophthalmus. Presence or absence of TLR4 and MyD88 in different tissues indicates functional importance of these tissues in innate immunity. Hence, tissue distribution of TLR4 and MyD88 was also judged by PCR method in this study. Tissue distribution analysis revealed presence of TLR4 in seven out of ten tissues screened. TLR4 was detected in blood, kidney, spleen, intestine, gills, brain and liver. However, TLR4 could not be detected in heart, skin and muscle tissues. MyD88 was detected in all ten tissues under study. Â-actin gene primers were used as internal control to confirm the quality of cDNA used in this study. The outcome of this research will be helpful for studying mRNA expression levels of TLR4 and MyD88 under various conditions in different tissues of P. hypophthalmus.Not Availabl