Next Generation Sequencing Facilitates Quantitative Analysis of Col, myb30 and MYB30-OE Transcriptomes

total mRNA profiles of 4-day-old Col (WT), MYB30 knockout mutant (myb30) and MYB30-OE were generated by deep sequencing, in triplicate, Sequencing was carried out with the Illumina HiSeq 2000 platform and the resulting reads were mapped to the reference genome of Arabidopsis thaliana (TAIR10) with TopHat (http://tophat.cbcb.umd.edu). Transcript expression was evaluated by cuffdiff (http://cufflinks.cbcb.umd.edu), and transcript abundance was estimated by fragments per kilobase of exon model per million mapped fragments (FPKM). Differentially expressed genes were selected using Student's t-test with P<0.05. qRT–PCR validation was performed using SYBR Green assays. Overall design: mRNA profiles of 4-day old Col, myb30 and MYB30-OE were generated by deep sequencing, in triplicate, using Illumina HiSeq 2000.