| Library |
| Library name |
Construction protocol |
Strategy |
Source |
Selection |
Layout |
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Crayfish were divided into three groups and were injected with WSSV-CN01, WSSV-CN03 (5×10^8 virions per individual), or an equal volume of normal saline. There were 15 animals in each group and the hemocytes from 5 animals were mixed as one sample. Thus, at each time point, there were 3 samples in each group. Hemocytes were collected from the animals at 0, 12, and 36 h post-infection. Total RNA was extracted from the cells using TRIzol (Invitrogen). After total RNA was extracted, eukaryotic mRNA was enriched by Oligo(dT) beads, while prokaryotic mRNA was enriched by removing rRNA by Ribo-ZeroTM Magnetic Kit (Epicentre). Then the enriched mRNA was fragmented into short fragments using fragmentation buffer and reverse transcripted into cDNA with random primers. Second-strand cDNA were synthesized by DNA polymerase I, RNase H, dNTP and buffer. Then the cDNA fragments were purified with QiaQuick PCR extraction kit, end repaired, poly(A) added, and ligated to Illumina sequencing adapters. The ligation products were size selected by agarose gel electrophoresis, PCR amplified, and sequenced using Illumina HiSeqTM 4000 by Gene Denovo Biotechnology Co. (Guangzhou, China). |
RNA-Seq |
TRANSCRIPTOMIC |
PCR |
PAIRED
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