siam betta fish ukIndels were initially established by manual alignment of genome sequences between homozygous mutant and wild type. Primers of these indels of appropriate length were then designed for PCR assays. Gene expression was studied by RT PCR. Total RNA from independent tissues or embryos was remoted using TRIzol reagent Invitrogen. Two micrograms of total RNA were treated with DNase I Roche after which used for cDNA synthesis using Reverse Transcriptase M MLV Promega. Expression of genes of interest in different tissues and in various developmental stages was initially examined using RT PCR with gene real primers.