Lateral root meristem establishment by LRIS in Arabidopsis
Lateral root meristem establishment by LRIS in Arabidopsis
root sections between 1000 µm and 2000 µm from the root tip were microdissected  Col-0 seeds were germinated on medium containing 10 μM NPA and transferred 3 d after germination under continuous light to 10 μM NAA, according to the points of the time course. total RNA was extracted using  Trizol (Invitrogen) and the RNeasy Plant mini kit (Qiagen) Using the Illumina TruSeq® Stranded mRNA Sample Prep Kit poly-A containing mRNA molecules were purified from the total RNA input using poly-T oligo-attached magnetic beads. In a reverse transcription reaction using random primers, RNA was converted into first strand cDNA and subsequently converted into double-stranded cDNA in a second strand cDNA synthesis reaction using DNA PolymeraseI and RNAse H. The cDNA fragments were extended with a single 'A' base to the 3' ends of the blunt-ended cDNA fragments after which indexing adapters were ligated introducing single distinct barcodes for each sample. Finally, enrichment PCR was carried out to enrich those DNA fragments that have adapter molecules on both ends and to amplify the amount of DNA in the library.