Efficient cDNA Preparation and Amplification of Laccase 2 gene from various tissues of Tasar Silkworm for Genetic Studies

Tasar silkworms, scientifically known as Antheraea mylitta, a species of wild silkworms native to India. The tasar silkworm undergoes a complete metamorphosis, including the egg, larva (caterpillar), pupa (cocoon), and adult (moth) stages. This complex life cycle, coupled with their outdoor rearing environment, makes their cultivation distinct and labour-intensive compared to other silkworm species. To improve the quality of silk being produced, it is of utmost importance that genetic analysis with preference to gene expression is well understood. Transcriptome analysis is one of the most crucial studies performed to understand the level of gene expression in a living organism. With respect to Antheraea mylitta, currently there are very limited genetic data describing the gene expressions. Transcriptome analysis at different stages of larvae development can give useful insight. The data can also be used to study the response of the insect's immune system in case of pathogen attack. The study focused on optimization of cDNA synthesis from RNA isolated from Antheraea mylitta. Our study used Laccase2 gene for PCR amplification to check the quality of isolated RNA. The study concluded that good quality RNA can be successfully isolated from Antheraea mylitta. Successful amplification of Laccase2 gene reveals that Laccase2 gene is being expressed at different levels of different larval stages as it helps in cuticle tanning and sclerotization. This research focuses on synthesizing cDNA for the Laccase 2 gene from Antheraea mylitta and confirming its identity using PCR amplification and sequence analysis, contributing to a deeper understanding of its biological function.