Methyl-MaxiSeq® libraries were prepared from 80 ng of genomic DNA digested with 2 units of Zymo Research’s (ZR) dsDNA Shearase™ Plus (Cat#: E2018-50).
Then, DNA from each sample was quantified and applied a QC check using the Qubit instrument according to the manufacturer’s guidelines (Qubit 3.0 Fluorometer Life Technologies – Carlsbad, CA, USA). There, samples were treated with RNAse (RNA removal) followed by clean-up using their genomic DNA Clean & Concentrator kit (Cat.
1 μg of DNA of each sample was resuspended in 50 μl of TE buffer to send them to Zymo Research (Irvine, CA) for the next steps in DNA methylation analysis (library construction protocol, sequencing run, data processing and data analysis). LiCl 2M was used for a first step of DNA purification, to remove RNA. of chloroform isoamyl (24:1 v:v) and for the precipitation was applied 1 vol. We applied 500 µl CTAB and 2 µl/ml β-mercaptoethanol, and samples were incubated 1h at 60 ✬. Each biological replicate for each condition were 4 plants.ĭNA was extracted from 100 mg of leaves powder from 3 biological replicates using CTAB extraction protocol. Seeds were previously stratified for 3 days. Treatment: Pseudomonas syringae DC3000 infiltrated (OD: 2x10^-4)Īrabidopsis thaliana plants were grown in jiffys in growth chambers (19-22✬, 10000 luxes fluorescent illumination) on a 10h-light and 14h-dark cycle for a month. GEO help: Mouse over screen elements for information.
0 kommentar(er)
