Transfer RNAs (tRNA) are important molecules that involved in protein translation machinery and acts as a bridge between the ribosome and codon of the mRNA. The study of tRNA is evolving considerably in the fields of bacteria, plants, and animals. However, detailed genomic study of the cyanobacterial tRNA is lacking. Therefore, we conducted a study of cyanobacterial tRNA from 61 species. Analysis revealed that; cyanobacteria contain thirty-six to seventy-eight tRNA gens per genome that encodes for 20 tRNA isotypes. The number of iso-acceptors (anti-codons) ranged from thirty-two to forty-three per genome. tRNA with anti-codon AAU, GAU, and UAU was reported to be absent from the genome of PCC 73,106 and sp. PCC 7305. Instead, they were contained anti-codon CAU that is common to tRNA and tRNA as well. The iso-acceptors ACA (tRNA), ACC (tRNA), AGA, ACU (tRNA), AAA (tRNA), AGG (tRNA), AAC (tRNA), GCG (tRNA), AUG (tRNA), and AUC (tRNA) were absent from the genome of cyanobacterial lineages studied so far. A few of the cyanobacterial species encode suppressor tRNAs, whereas none of the species were found to encode a selenocysteine iso-acceptor. Cyanobacterial species encode a few putative novel tRNAs whose functions are yet to be elucidated.