GSM1260853: CS; Manihot esculenta; RNA-Seq
Three organs/tissues (folded leaf, fully expanded leaf and roots) of Cassava cultivar SC124 harvested at 6h, 24h and 5d for three cold treatments of CA, CCA and CS, for gene expression profiling at the stages of initial response, secondary response, and functional adaption to cold stresses. Total RNA of each sample was isolated individually, and then pooled with an equal amount from each sample into one for profiling. As a result, four mRNA libraries and four small-RNA libraries, corresponding to the conditions of CA, CCA, CS and NC, were constructed. The four mRNA libraries were sequenced by RNA-seq by Illumina GAII following Illumina RNA-seq protocol. Briefly, total RNAs were isolated, purified and reversely transcribed, the resulting cDNA products were subsequently digested with NlaIII and the 3′-cDNA fragments captured with the oligo(dT) beads, and then ligated to the Illumina GEX NlaIII Adapter 1. The junction of Illumina adapter 1 and CATG site contained the recognition site of MmeI, which cutting 17 bp downstream of the recognition site (CATG) to produce tags. After removing 3′fragments with magnetic beads precipitation and MmeI digestion, an Illumina GEX adapter 2 was introduced at the end of tags. The resulting adapter-ligated cDNA tags were subjected to 15 cycles of linear PCR amplified, purified and sequenced with the method of sequencing by synthesis (SBS) using Illumina GAII
GEO Accession
GSM1260853