Affymetrix GeneChip Arabidopsis Genome [ATH1-121501]

Common transcriptional responses of Arabidopsis thaliana protoplasts transfected with TCV, HCRSV and their coat protein mutants

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Common transcriptional responses of Arabidopsis thaliana protoplasts transfected with turnip crinkle virus (TCV) , hibiscus chlorotic ringspot virus (HCRSV) and their coat protein mutants.

plant viral treatments

Transcription profiling of Arabidopsis protoplasts transfected with TCV or HCRSV and their coat protein mutants

Common transcriptional responses of Arabidopsis thaliana protoplasts transfected with turnip crinkle virus (TCV) , hibiscus chlorotic ringspot virus (HCRSV) and their coat protein mutants.


Affymetrix:Protocol:ExpressionStat

Title: Affymetrix CHP Analysis (ExpressionStat). Description:

Affymetrix:Protocol:Hybridization-EukGE-WS2v4[]

Title: Affymetrix EukGE-WS2v4 Hybridization. Description:

P-AFFY-2

Labeling of samples was carried out according to the manufacturer’s standard protocol

P-AFFY-6

Title: Affymetrix CEL analysis. Description:

P-MEXP-82989

Arabidopsis thaliana ecotype Columbia 3 (Col-3) protoplasts was chosen for this study. The plants were grown under conditions of 12 h light/dark at 25 oC. Fully expanded leaves were harvested from 3-4 weeks old seedlings and surface sterilized for 10 min with 0.8 % Clorox® (sodium hypochloride), followed by 5 min washing with MilliRO® water for three times. The leaves were sliced into 1 mm strips and agitated in enzyme mixture slowly for 3 h at 25 °C in the dark to isolate the protoplasts. 

P-MEXP-82990

The protoplasts were kept on ice during all isolation steps and centrifugation was conducted at 4 °C. Protoplasts of 200 ìl were mixed gently with 20 ìg RNA transcripts (TCV, HCRSV, TCV-HCP, HCRSV-TCP) and 200 ìl polyethylene glycol (PEG) solution (40 % PEG, 0.3 M mannitol, 100 mM CaCl2). Immediately after the mixing, 4.5 ml W5 solution was added to dilute the protoplasts. The protoplasts were kept on ice for 20 min before centrifugation at 100 x g for 3 min. Supernatant was discarded and the protoplasts were washed again. Protoplasts were incubated in protoplasts culture medium in a Petri-dish at a concentration of 1 X10 power of 5 at 25 °C in the dark for 16 and 48 hr respectively.

P-MEXP-82994

Total RNA was extracted from protoplasts at 16 h post transfection using RNeasy Mini Kit (Qiagen, Germany) according to manufacturer’s instructions.

Transcription profiling of Arabidopsis protoplasts transfected with TCV or HCRSV and their coat protein mutants

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