Team:Paris Saclay/Notebook/June/28
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Contents
Tuesday 28th June
Lab work
Interlab study
Cytometer
"By Caroline, Alice, Lea and Charlene, with Isabelle help
Cells culture created on the 27/06/2016 were analysed with cytometer. Results were as expected.
Visualization
Puc19 digestion
Puc19 was digested as expected.
Puc19-gBlocks ligation
By Charlene
GBlocks were resuspended into 100µL of TE buffer (except for 4-1 which was resuspended into 50µL of TE) in order to get a concentration of 10ng/mL after a quick spin. Solutions were vortexed, incubated for 20min at 50°C, vortexed another time and quickly spun down.
| gBlock | 1-1 | 1-2 | 2-1 | 3-1 | 3-2 | 4-1 | 4-2 | GFP1-9 |
|---|---|---|---|---|---|---|---|---|
| Size (bp) | 960 | 960 | 1023 | 960 | 960 | 706 | 1288 | 862 |
| vector size/insert size x 3.5 | 9.84 | 9.84 | 9.24 | 9.84 | 9.84 | 13.39 | 7.34 | 10.96 |
3 controls were made :
- 1µL digested vector + 9µL H2O
- 1µL digested vector + 1µL ligase + 1µL ligation buffer + 7µL H2O
- 1µL digested vector + 1µL ligase + 1µL ligation buffer + 7µL insert
Transformation with ligation products
By Caroline and Charlene
DH5α cells were transformed with ligation products using the same protocol as on 24/06/2016 Transformations were displayed on Petri dishes with LB medium containing 50µg/mL of ampicillin and covered with 0.5µL of 80mg/mL XGal and 0.5µL of 1M IPTG.
Bringing DNA closer
Plasmid extraction
By Naiane and Lea
Plasmids from cell culture of the 27/06/2016 were extracted following the same protocol as on 24/06/2016.
Cells transformation
By Naiane
Extracted plasmids (2µL) were transformed into DH5α competent cells (50µL) using the same protocol as on 24/06/2016.
BioBrick characterization
Plasmid extraction
By Lea
The same protocol as on 24/06/2016 was used to extract K1372001 from DH5α cells, except the phenol chloroform step was not done.
