Header

1) Third PCRs (50µl)
- For IYS: PCR (5) to add Gibson overhangs on BBa_K523013_PCR_product (3) where linker were added the day before
- For ISY: PCR (6) to add Gibson overhangs on BBa_K523013_PCR_product (4) where linker were added the day before
- PCR purification

2) Gibson Assembly
- IYS Gibson Assembly using PCR (1) from the day before and PCR (5)
- ISY Gibson Assembly using PCR (2) from the day before and PCR (6)

- 0.8% Gel electrophoresis to verify the thirs PCRs and the Gibson Products => the products are higher than the expected sizes but we decided to do the transformation anyway.

2)DH5_alpha Transformation
- Transformation of the two Gibson products with 2 controls consisting of the backbone and the insert use for the Gibson (PCR (1) & (5) for IYS and PCR (2) & (6) for ISY)
- Additionnal controls => competent cells and BBa_K523013 miniprep (initial template used for the PCRs)
- cells were plated on Chloramphenicol plates and grow overnight in a 37°C incubator