AT0075: 4x18:6-DD

Property Value
BioDare ID 12729849314623
Author Adrian Thomson
Institution University of Edinburgh
License CC_BY

Description

LUC imaging 18L:6D into constant dark

Purpose

testing phase changes of clock components under different photoperiods

Comments

microwave seed sterilisation Pedro prepared by editing groups from previous template

Sample preparation

sprayed upon transfer from entrainment in Sanyo chamber to Percival cabinet

Measurement

Protocol

Luciferase luminescence (HAM 3)

Standard Adrian Thomson protocol for imaging

imaging script starts 5 minutes before required ZT, then 55 minutes interval, 5 minutes light off, 30 minutes capture

Experimental conditions

LD18:06 22c

Growth on Agar 1.5% MS 1 3% Sucrose for 6.0 days (LD18:06 22c). entrainment

Light

Type Duration (days) Cycle (h) Start Duration Spectrum Source Intensity
diurnal light 6 24 0:00 18:00 white tube 100

Temperature

Type Duration (days) Cycle (h) Base (°C) Warm (°C) Warm start Warm duration
constant temperature 6 24 22 -- -- --

4x18:6-DD

Growth on Agar 1.5% MS 1 3% Sucrose for 7.0 days (4x18:6-DD). first day not imaging, 4 days 18L:6D, 3 days constant dark P14H3 (Percival chamber number 14; Hamamatsu camera 3)

Light

Type Duration (days) Cycle (h) Start Duration Spectrum Source Intensity
diurnal light 4 24 0:00 18:00 blue LED 10
constant dark 3 24 -- -- blue LED 10
diurnal light 4 24 0:00 18:00 red LED 10
constant dark 3 24 -- -- red LED 10

Temperature

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

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