GSM9203910: tph_1_CAM788_6; Caenorhabditis elegans; RNA-Seq
Fresh working lysis buffer is prepared daily by mixing a stable lysis buffer stock (made of Tris-HCl, Triton-X, Tween-20, EDTA, and water), Proteinase K, and RNase inhibitor. Worms were collected individually, and cut with a needle if larval. After adding lysis buffer, samples are mixed, centrifuged, and incubated in a thermocycler at 65 °C for 10 min, 85 °C for 1 min, then 4 °C before cooling on ice. Library preparation for each single individual was according to the Illumina Nextera tagmentation protocol (Illumina), with DNA input and reagents adjusted for 20 ng of cDNA. The tagmented DNA was cleaned, and PCR amplification was performed. PCR primers were removed using Ampure XP beads.