The study was designed to test the hypothesis, variations in nucleotide frequency distribution in cytochrome c oxidase subunit I (COI) in spiders used as a marker to delimit species. Based on morphological identification and sequencing of CO1 gene, a total of 22 species of spiders belonging to five families were investigated and compared within each species and among families for nucleotide frequency distribution of all species collected. We had found non-significant difference in frequency of nucleotides in COI within the members of species and within families (thymine 41.30-44.30%, adenine 24.96-27.19%, guanine 17.18-19.48% and cytosine 12.28-13.40% while GC 29.53-32.84%). Therefore, it was concluded that variation in nucleotide frequency distribution could not to bee used as a criterion for species delimitation in spiders.
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