Dataset features


Application: RNA-seq analysis
Number of samples: 2
Release date: Nov 8 2016
Last update date: Aug 14 2018
Access: Public
Diseases: Deficiency Diseases, Citrullinemia
Genes: FPS1, FPS2
Dataset link FPS down-regulation alters chloroplast development and triggers an sterol-dependent induction of JA-related and Fe deficiency transcriptional responses

Experimental Protocol

To induce FPS gene silencing, seedlings were germinated and grown for three days on sterile filter papers that were first placed on MS plates and subsequently transferred onto new MS plates supplemented (treatment) or not (control) with 30 µM MFZ for five days more. Three independent pools of seedlings (100 mg of FW) were collected per each treatment. Seedlings were ground to a fine powder using TissueLyser equipment and used for extraction of RNA. The quality and quantity of total RNA samples were assessed using a Bionalyzer Expert 2100 Instrument (Agilent Technologies) and an equimolar mixture of RNA samples from each treatment was prepared. The RNA samples (3 μg) were fragmented and ligated with adaptors prior to cDNA synthesis and PCR amplification. The cDNA libraries were prepared according to the Illumina protocols and sequenced using an Illumina HiSeq2000 machine to perform 2x100 paired-end sequencing.










David Manzano