GSM3552950: BSPCR_Hairpin_x_ZF_line2; Arabidopsis thaliana; OTHER
Leaf tissue from adult plants of representative T2 lines was collected to analyze FWA methylation. Inflorescences of representative T2 lines were collected to analyze AP1 methylation. DNA was extracted following a CTAB-based method and converted using the EZ DNA methylation-lighting kit (ZYMO research) or the Epitect Bisulfite Conversion kit (Qiagen). For FWA, we analyzed methylation over three different regions: Region 1: chr4: 13038143-13038272; Region 2: chr4: 13038356-13038499; Region3: chr4: 13038568-13038695. For AP1, we analyzed methylation over three different regions: Region 1: chr1: 25986270-25986391; Region 2: chr1: 25985976-25986098; Region 3: chr1:25986235-25986108. To amplify the different regions, Pfu Turbo Cx (Agilent) was used together with primers containing the Illumina adaptors. The primers used are listed in Table S1. Different PCR products for the same sample were pooled and purified by AMPure beads (Beckman Coulter). Libraries were made from purified PCR products by a TruSeq Nano DNA Library Prep kit for Neoprep automated library preparation machine (Illumina), by Kapa DNA hyper kit (Kapa Biosystems) with Illumina TruSeq DNA adapters or by Ovation Ultralow V2 kit (NuGEN).
GEO Accession
GSM3552950