Team:Bielefeld-CeBiTec/Notebook/HeavyMetals
- We transformed BioBricks from the distribution:
- BBa_J33201 (ArsR)
- BBa_K516030 (mRFP1 with RBS and terminator)
- Colonies were used to inoculate overnight cultures.
- Plasmids were isolated using a miniprep kit.
- ArsR (BBa_J33201) and RFP (BBa_K516030) were combined using 3A assembly:
- Restriction digest of arsR, RFP and pSB1K3.m1
- PCR Clean-up
- Ligation: 2 µL per fragment, 30 min
- Transformation by electroporation and heat shock (for comparison)
- No colonies, therefore the ligation was repeated with significantly more DNA. Incubation over night.
- Electroporation resulted in colonies, which were screened with a colony PCR.
- A product length of 1705 bp was expected. The gel shows that most plasmids seem to be correct.