RNA-Seq of C.elegans: flr-4(n2259)Day1_01
L1 synchronized worms flr-4(n2259) were grown on OP50 till L4/YA, after which FUDR was added to prevent progeny growth. After about 24 hours, worms were collected in M9 buffer and washed thrice and then frozen in Trizol (Invitrogen, USA). The worms were collected, passed through a freeze-thaw cycle and lysed by vigorous vortexing. RNA was purified by phenol:chloroform:isoamylalcohol extraction followed by ethanol precipitation. RNA Quality Control of RNA was done using Bioanalyzer 2100 RNA 6000 NanoAssay chip and RNA above RIN = 8 was included to the study (Agilent Technologies, Santa Clara, USA).  cDNA library was subsequently constructed by TruSeq RNA Library Prep Kit v2 (Illumina). Sequencing (76 bp single end) was performed with two biological replicates using Illumina Genome Analyzer IIx (GA IIx). Imaging, base calling and quality scoring were performed as per standard manufacturers guidelines (Illumina, USA).The de-multiplexing and conversion of BCL file format reads to FASTQ file format was done with Illumina-supported CASAVA v1.8.2 software package.