GSM1686457: 6dpf mpeg1:mCherryPosMmarinumGFPPos_rep1; Danio rerio; RNA-Seq
Between 200 and 300 larvae per condition were collected at 6 dpf. Larvae were dissociated into single cell suspension by Trypsin treatment and fluorescent and 20 fluorescent cells per condition were sorted by FACS directly into cDNA synthesis buffer, without RNA extraction. cDNA were synthetized and amplified with the SMARTer cDNA synthesis kit for Illumina Sequencing (Clonetech) according to the manufacturer’s instructions. The integrity of the cDNA was confirmed by Lab-on-chip analysis using the 2100 Bioanalyzer (Agilent Technologies). The sheared amplified cDNA  were used to make RNAseq libraries using the Illumina TruSeq RNA Sample Preparation Kit v2 (Illumina Inc., San Diego, USA). In the manufacturer’s instructions two modifications were made. In the adapter ligation step 1 µl instead of 2.5 µl adapter was used. In the library size selection step the library fragments were isolated with a double Ampure XP purification with a 0.7x beads to library ratio. The resulting mRNA-Seq library was sequenced using an Illumina HiSeq2000 instrument according to the manufacturer’s description with a read length of 1 x 50 nucleotides.
GEO Accession
GSM1686457