title: GSM3892304: RagA_mutant_rep1; Caenorhabditis elegans; RNA-Seq
library_construction_protocol: Aproximately 1000 mid-L4 stage animals were washed, resuspended in QIAzol lysis reagent (Qiagen, Venlo, Netherlands, #79306), snap frozen in liquid nitrogen, and stored in -80°C freezer until RNA extraction. To break the worm cuticle and improve RNA yield, all samples underwent 5 freeze-thaw cycles. In each cycle, samples were thawed at 37˚C and then snap-frozen in liquid nitrogen. RNA extraction was performed immediately using QIAGEN RNeasy Mini Kit (QIAGEN, #74104) following manufacturer's instructions. RNA quality was confirmed using Agilent Bioanalyzer 2100 (Agilent Technologies). All samples passed the quality control standard of RIN>8.0. mRNA libraries were prepared using TruSeq® Stranded mRNA LT - Set A kit (Illumina, CA, USA RS-122-2101) following manufacturer's instructions and linked to different adaptors to enable pooling. Library quality was checked using 2200 High Sensitivity D1000 Tape Station (Agilent Technologies) RNA-Seq with Illumina HiSeq 2500 using 50-cycle, pair-end settings
attributes.tag: GEO Accession
attributes.value: GSM3892304