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Contents
Tuesday 28th June
Lab work
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