"

From 2014.igem.org

We picked up more stabs from iGEM HQ. We got three new chromoproteins: BBa_K1073023 (eforRed [pink]), BBa_K1033931 (amilGFP [yellow]), and BBa_K1033929 (aeBlue [indigo]). While we were there, we also got a plate with BBa_I765001 (UV promoter) and BBa_K274200 (orange pigment) because we originally had trouble growing them up (we thought they were in a different backbone, and therefore grew them up in the wrong antibiotic).

PCR Results (Red pigment, BLux, RLux, HSP, asp) for Gibson reaction: Ran reactions on gel: Many failed reactions (for the Blux inserts, and HSP & ASP insert and backbones):

We then decided to run the failed PCR reactions at a range of annealing temperatures 50C-75C (See PCR Planning spreadsheet for specifics) to troubleshoot the problem.

We gel purified the PCRs that worked (Red pigment and Blux), set up the Gibson reactions, transformed, and plated them.

We sequenced the Gibsoned hybB promoter in pBbE1a construct (pHBD17), the chromoproteins (pHBD32,33,34) and the SB3K3 backbone (HBD35).

We also made more CM and Crb plates.

We grew up BBa_K274200, BBa_I765001, pHBD4, BBa_K1073023, BBa_K1033931, and BBa_K1033929 to miniprep tomorrow.