De novo tissue specific transcriptome assembly by using short reads of cDNA samples from various tissues

Genomic and transcriptomic data related to Astacus leptodactylus is fairly limited. We have endeavored to assemble tissue specific transcriptomes by using large amount of (illumina) short read clusters for each tissue of interest. Total RNA was extracted from ganglia, muscle, and receptor organ samples individually. A conventional phenol-chloroform method was used for extracting RNA from the tissue of interest. Concentration of the RNA extract was measured by a florometric device (Qubit; Thermo Fisher Scientific, Waltham, MA, USA). RNA samples were reverse-transcribed into DNA copies by using a transcriptase in the presence of random and oligo dT primers. Synthesized DNA was ligated by using a high-efficiency ligation mix. The ligated DNA was amplified by using SensiPhi DNA polymerase in an isothermal reaction for 2 h (Repli-g WTA kit; Qiagen, Hilden, Germany). The final concentration of the double stranded cDNA library was about 0.3-0.5 ug ul.