Q-PCR LD (12:12) 17C LHY

Property Value
BioDare ID 3492
Author Aurora Pinas-Fernandez
Institution University of Edinburgh
License CC_BY

Description

Literature data from: 'Temp' by: Aurora Pinas-Fernandez.

    Wild-type and clock-mutant seedlings were entrained for 7 days at 22C in 12:12 white light. Then shifted to 17C 12:12 white light. Samples were collected on the 6th day at 17 degrees, every 2h, for 24h.

Purpose

Experiment to reparameterize the clock model without exogenous sucrose

Comments

24h sample lost for several genotypes. Curated by Andrew Millar. AJM manually edited the 14h to 18h timepoints in the prr7-9- mutant, as many values were omitted when the ratio's looked reasonably close among reps. For toc1-9, ignore LHY data, much lower than CCA1 in first 4 timepoints. These datapoints removed from file.

Sample preparation

Samples were harvested into Ambion RNAlater for some weeks, at 4C and -20C. RNA extraction was done following protocol of GE Health Care-illustra RNAspin 96RNA Isolation kit. Samples were spec'd with Nanodrop. cDNA synthesis was done with Super Script Vilo kit (Invitrogene) from 1ug RNA.

Measurement

Protocol

qRT-PCR (Robot + Light Cycler 480)

Literature data from: '' by: .

    primers unknown

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Experimental conditions

Entrainment 22C Diurnal

Growth on Agar 1.2% MS 0.5 for 7.0 days (Entrainment 22C Diurnal).

Light

Type Duration (days) Cycle (h) Start Duration Spectrum Source Intensity
diurnal light 7 24 0:00 12:00 white tube 50

Temperature

Type Duration (days) Cycle (h) Base (°C) Warm (°C) Warm start Warm duration
constant temperature 7 24 22 -- -- --

17C LD (12:12) Off Sucrose

Growth on Agar 1.2% MS 0.5 for 6.0 days (17C LD (12:12) Off Sucrose).

Light

Type Duration (days) Cycle (h) Start Duration Spectrum Source Intensity
diurnal light 6 24 0:00 12:00 white tube 50

Temperature

Type Duration (days) Cycle (h) Base (°C) Warm (°C) Warm start Warm duration
constant temperature 6 24 17 -- -- --

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