Team:Paris Saclay/Notebook/June/28

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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 :

  1. 1µL digested vector + 9µL H2O
  2. 1µL digested vector + 1µL ligase + 1µL ligation buffer + 7µL H2O
  3. 1µL digested vector + 1µL ligase + 1µL ligation buffer + 7µL insert

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