title: Illumina HiSeq 2500 paired end sequencing; Thioredoxin DCC1 regulates shoot regeneration through modulating multiple genes expression by RNA-seq analyses in Arabidopsis
design: Thioredoxin DCC1 regulates shoot regeneration through modulating multiple genes expression by RNA-seq analyses in Arabidopsis
library_construction_protocol: Three biological repeats of the wild type (Col-0-A, Col-0-B and Col-0-C) and the mutant dcc1 (dcc1-A, dcc1-B and dcc1-C) after culture on shoot-induction medium for 16 days were used for RNA sequencing. Total RNAs were isolated from the calli of wild type Col-0 and the mutant dcc1 with TRI reagent (Sigma, catalog number T9424). The mRNA was enriched by Ribo-ZeroTM Magnetic Kit (Epicentre), and fragmented into short fragments using fragmentation buffer and reverse transcripted into cDNA with random primers. Second-strand cDNA were synthesized by DNA polymerase I. 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, and amplified by PCR to construct the RNA-seq llibraries.
attributes.tag: Experimental Factor: wild type genotype
attributes.value: genotype