| Property | Value |
|---|---|
| BioDare ID | 13982031569136 |
| Author | Laura Dixon |
| Institution | University of Edinburgh |
| License | CC_BY |
pTOC plate 0683 pCAB plate0684 LHY plate 0685 control plate 0686 TOC plate0687 pLHY plate 0688
Re-entrainment dynamics during the transition between light:dark cycles with different photoperiods, comparing Arabidopsis thaliana and Ostreococcus tauri. This expt in O. tauri compares transcriptional LUC fusions to TOC1, CAB, and LHY promoters with translational fusions to LHY and TOC promoter + protein. Each marker is on a separate plate to avoid plate-position effects.
Several controls are relevant, from parallel experiments in adjacent weeks. Data displayed in BioDare are means of 8 cultures in a 96-well plate column, where data from each culture was normalised to its mean before averaging. 6 such columns were measured for each marker (each marker has its own plate). Raw data are shown in the 'normalised' files. Aggregated data in the 'summary.xls' file. NB each plate is counted in sequence, so necessarily at a different time. The four LHY/TOC1 markers are counted within 0.5h, in a total cycle time of 0.707h. For display in BioDare, the times are shown for the middle plate, TOC, but the separate files have the original, separate times. The peak analysis in the publication used the averages for each column.
Cultures were in alternate columns of the 96-well plates (i.e. columns 1, 3, 5, ... 11) to avoid light crosstalk. This also speeds up plate reading in this two-PMT instrument, which reads columns 1 and 7 together, then skips 2 and 8, etc. The artificial seawater was refreshed and luciferin was applied 24 h before assays started.
Luciferase luminescence (Top count1)
Control plate shows background <150 cps in L, <30cps in D, insignificant, so no background subtracted.
Growth on Artificial Sea Water (ASW) with Keller supplements, plus antibiotics for 7.0 days (8L:16D). White fluorescent tubes filtered with one layer of Ocean Blue gel filter. NB fluence rate was 10 micromol/m2/s. Earlier papers stated 20, this was incorrect.
| Type | Duration (days) | Cycle (h) | Start | Duration | Spectrum | Source | Intensity |
|---|---|---|---|---|---|---|---|
| diurnal light | 7 | 24 | 0:00 | 8:00 | white | tube | 10 |
| Type | Duration (days) | Cycle (h) | Base (°C) | Warm (°C) | Warm start | Warm duration |
|---|---|---|---|---|---|---|
| constant temperature | 7 | 24 | 20 | -- | -- | -- |
Growth on Artificial Sea Water (ASW) with Keller supplements, plus antibiotics for 7.0 days (SD to LD). In the stacker of the TopCount with red and blue LEDs
| Type | Duration (days) | Cycle (h) | Start | Duration | Spectrum | Source | Intensity |
|---|---|---|---|---|---|---|---|
| diurnal light | 2 | 24 | 0:00 | 8:00 | red | LED | 5 |
| diurnal light | 5 | 24 | 0:00 | 16:00 | red | LED | 5 |
| diurnal light | 2 | 24 | 0:00 | 8:00 | blue | LED | 5 |
| diurnal light | 5 | 24 | 0:00 | 16:00 | blue | LED | 5 |
| Type | Duration (days) | Cycle (h) | Base (°C) | Warm (°C) | Warm start | Warm duration |
|---|---|---|---|---|---|---|
| constant temperature | 7 | 24 | 20 | -- | -- | -- |