Difference between revisions of "Team:Reading/Notebook"
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<li>SODIS 3 - 1ml of stock Howe Lab triple mutant 6803.</li></ul></li> | <li>SODIS 3 - 1ml of stock Howe Lab triple mutant 6803.</li></ul></li> | ||
</ol> | </ol> | ||
− | <p> All flasks cultured today were placed in the incubator, being shaken at 58rpm, 30°C, and under bright white light.</p> | + | <p> All flasks cultured today were placed in the incubator, being shaken at 58rpm, 30°C, and under bright white light </p> |
+ | <p> <b> 05/08/15 </b> </p> | ||
+ | <p> 200ml of BG-11 was added to flasks 3 and 4 which already contained ~100ml of ~2 a.u density 6803 WT. </p> | ||
+ | <p> Competition Assay </p> | ||
+ | <p> 200ml of BG-11 added to a new sterile conical flask </p> | ||
+ | <p> added 10ul of WT 6803 and 10ul of MT3 6803 </p> | ||
+ | <p> Flask was placed in the incubator, being shaken at 58rpm, 30°C, and under bright white light </p> | ||
+ | <p> <b> 06/08/15 </b> </p> | ||
+ | <p> DNA concentration from miniprep </p> | ||
+ | <ul> | ||
+ | <li>PilA1 = 11.16ns/ul </li> | ||
+ | <li>PsaD = 13.01ns/ul </li> | ||
+ | <li>PetF = 0.38ns/ul </li> | ||
+ | <li>PilT = 8.58ns/ul </li> | ||
+ | </ul> | ||
+ | <p> *no DNA due to miniprep from Glycerol Stock </p> | ||
+ | <p> <b> 07/08/15 </b> </p> | ||
+ | <p> Made LB Agar tranplates for culturing </p> | ||
+ | <ul> | ||
+ | <li>PilA1</li> | ||
+ | <li>PsaD</li> | ||
+ | <li>PetF</li> | ||
+ | <li>PilT</li> | ||
+ | </ul> | ||
+ | <p> <b> 10/08/15 </b> </p> | ||
+ | <p> No growth on plates from 07/08 </p> | ||
+ | <p> Cultured plates of LB Agar from all stocks (glycerol stocks) </p> | ||
+ | <p> <b> 11/08/15 </b> </p> | ||
+ | <p> Growth on many the stocks from 10/08, replated onto LB kan 50 plates </p> | ||
+ | <p> <b> 12/08/15 </b> </p> | ||
+ | <p> Competition Assay had began to turn a light green colour. </p> | ||
</div> | </div> | ||
</html> | </html> |
Revision as of 13:03, 24 August 2015
This page is an account of the daily events in our lab and project. Dates are in the UK format DD/MM/YY. 6803 refers to the cyanobacterium used - Synechocystis sp. PCC 6803
29/06/15
All placed on the windowsill in our lab under natural light and at room temperature (~25°C)
02/07/15
07/07/15
10/07/15
2 Litres of water collected from Whiteknights Lake on campus for SODIS, method outlines in protocols.
13/07/15
Estimating the Cells per ml for the flask from 29/06 1 a.u at 750nm = 1.6x108
OD750 measurement and cells per ml estimated for flasks cultured on 02/07
14/07/15
The lamp safety testing completed so flasks placed under the bright white light bulb
21/07/15
Flasks are getting very green, definitely a good sign!
23/07/15
Flask 2 discarded as it is not showing growth, and has signs of contamination. We received most of the lab equipment we have ordered.
27/07/15
BG-11 order arrived finally!Miniprep of plasmid DNA: We carried out extraction of biobrick construct plasmids from E.coli NEB-5α glycerol stocks.
We extracted pSB1K3 plasmid backbones combined with biobricks containing the following genes:
- PilA1
- PsaD
- PetF
- PilT
This gave us 4 samples of plasmid DNA, which were placed in the freezer at -20°C.
-
BG-11 cultures: We added ~200ml of BG-11 to two sterile 500ml conical flasks. We inoculated them as follows;
- Flask 5 - 1ml of Wild type 6803 from flask 3 as this flask had the best growth.
- Flask 6 - 10µl of stock Howe Lab triple mutant 6803.
- SODIS lakewater cultures: We added ~200ml of SODIS lakewater to three sterile 500ml conical flasks. We inoculated them as follows;
- SODIS 1 and 2 - 1ml of Wild type 6803 from flask 3 as this flask had the best growth.
- SODIS 3 - 1ml of stock Howe Lab triple mutant 6803.
All flasks cultured today were placed in the incubator, being shaken at 58rpm, 30°C, and under bright white light
05/08/15
200ml of BG-11 was added to flasks 3 and 4 which already contained ~100ml of ~2 a.u density 6803 WT.
Competition Assay
200ml of BG-11 added to a new sterile conical flask
added 10ul of WT 6803 and 10ul of MT3 6803
Flask was placed in the incubator, being shaken at 58rpm, 30°C, and under bright white light
06/08/15
DNA concentration from miniprep
- PilA1 = 11.16ns/ul
- PsaD = 13.01ns/ul
- PetF = 0.38ns/ul
- PilT = 8.58ns/ul
*no DNA due to miniprep from Glycerol Stock
07/08/15
Made LB Agar tranplates for culturing
- PilA1
- PsaD
- PetF
- PilT
10/08/15
No growth on plates from 07/08
Cultured plates of LB Agar from all stocks (glycerol stocks)
11/08/15
Growth on many the stocks from 10/08, replated onto LB kan 50 plates
12/08/15
Competition Assay had began to turn a light green colour.