2024-05-16
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA114501744
External Id
Université Paris-Saclay, INRAE, AgroParisTech, Institut Jean-Pierre Bourgin (IJPB), 78000, Versailles, France.
INSDC center name
2023-10-16T18:18:33Z
INSDC last update
public
INSDC status
ERS16483396
SRA accession
E-MTAB-13442:MeioMeNGS_3
Submitter Id
ArrayExpress
broker name
not collected
collection date
thale cress
common name
Col-0
cultivar
Protocols: Liquid nitrogen frozen 15 days-old in vitro culture plants. Arabidopsis seeds were surface sterilized for 10 min in 70% ethanol + 0.05% sodium dodecyl sulfate and washed for 10 min in 70% ethanol and grown in Petri dishes with culture medium (Gamborg B5 medium-Duchefa supplied with 0.16% bromocresol purple and 0.1% sucrose). To generate the RFP-tagged NUP54, the NUP54 (AT1G24310) genomic fragment was amplified with NUP54_GTWU (XXX) and NUP54_GTWL (XXX) primers. The amplification covered NUP54 from the 990 pb before ATG to the last pb before the stop codon. The PCR product was cloned into pDONR207 to create pENTR-NUP54. For plant transformation, the LR reaction was performed with the binary vector pGWB553 (Nakagawa et al., 2007). The pGWB553-NUP54 was introduced into the Agrobacterium tumefaciens strains C58C1. Arabidopsis transformation was performed using the floral dip method: Agrobacterium tumefaciens was grown at 28–30 °C to saturation, centrifuged, and resuspended in a 1% sucrose 0.05% Silwet l-77 solution. This solution is used for dipping A. thaliana plants. Plants were maintained under glass overnight to increase humidity (Clough and Bent, 1998). For selection of transformants, sterilized seeds were sown on culture medium with 25 µg/mL hygromycin B (Duchefa). The generation of RFP-tagged REC8 (REC8 :RFP) line in (Kuttig et al., 2022) DNA was prepared using the NucleoSpin Plant II kit (Macherey Nagel) following the manufacturer’s instructions. Standard genomic library construction
description
rosette growth stage
developmental stage
transformant with CENH3 genomic fused with GFP
genotype
not collected
geographic location (country and/or sea)
not applicable
isolate
Arabidopsis thaliana
organism
Arabidopsis thaliana
organism
rosette leaf
organism part
Arabidopsis thaliana
scientific_name
MeioMeNGS_3
title