ES-2018-3
the raw data of crab hepatopancreas transcriptome
The hepatopancreases of crabs in the control, N1 and N3 groups were chosen for the RNA-seq analysis. Total RNA of hepatopancreases was extracted using TRIzol Reagent according to the manufacturer's instructions (Invitrogen), and the extracted RNA was treated with DNase I (Takara, Japan) to remove genomic DNA. The quality and quantity of total RNA were assessed using a Nano Drop 2000 spectrophotometer (Thermo, Wilmington, DE, USA).The RNA-seq transcriptome library was prepared following the TruSeq RNA sample preparation kit from Illumina (San Diego, CA) using 1g of total RNA. mRNA was isolated according to the poly A selection method using Oligo (dT) beads and then was fragmented using fragmentation buffer. Single-stranded cDNA was synthesized with random hexamers using RNA as template. Double-stranded cDNA was synthesized with the effect of dNTPs, DNA polymerase I, RNase H and buffer, and it was purified by AMPure XP beads. A single (A) was added using Klenow buffer.  Adaptor-modified fragments were selected by AMPure XP beads, and PCR amplification was performed for 15 cycles. After being quantified by Qubit 2.0, the sequencing library was diluted to 1.5ng/ul. The insert size of the library was tested by Agilent 2100, and was quantified quantitatively by Q-PCR method to guarantee the quality of the sequencing library. RNA-seq sequencing library was sequenced using Illumina HiSeq 4000.
ES-2018-3
SUB3920312
ES-2018-3