Team:LambertGA/Notebook

Home

Team

Team Collaborations

Project

Description Design Experiments Proof of Concept Results Notebook

Parts

Parts Basic Parts Composite Parts

AttributionsSafety

Human Practices

Human Practices Silver Gold Integrated Practices Engagement

Awards

Hardware Measurement Model

Notebook

Week 1 (Aug 8):
As a part of our biotechnology pathway’s curriculum, we reviewed basic safety procedures and aseptic technique with our advisor Janet Standeven.

Week 2 (Aug 15):
Our focus this week was to ligate two of the three parts in our genetic construct: P-lambda-R--LacI--TS Purple--LAA/DAS and R0011--ClpXP--CI; however, we did not ligate the part without any degradation tag. After the ligations, the constructs were transformed into DH10 E. coli cells, but we were unsuccessful.

Week 3 (Aug 22):
Due to the unsuccessful transformations seen the previous week, we re-attempted our workflow (digest, ligation, transformation) of both parts. In addition, we ligated and transformed our third construct: P-lambda-R--LacI--TS Purple and R0011--ClpXP--CI.

Week 4 (Aug 29):
Transformations were ultimately unsuccessful because of RFP (red fluorescent protein) contamination. In addition, we miniprepped and nanodropped our genetic construct without the present degradation tag, which was in the 1C3 backbone.

Week 5 (Sep 5):
Due to several failures over the past few weeks, we attempted to detect previous errors by re-ligating and re-transforming the parts together using a different stock of P-lambda-R--LacI that was previously confirmed over the summer. Our results indicated faint, purple cells for our construct (P-lambda-R--LacI--TS Purple--R0011--ClpXP--CI), but no color was shown for the DAS and LAA cells after the transformation. In addition, we inoculated liquid cultures of our genetic constructs from the previous week’s transformations, including the genetic construct without the present degradation tag, the RFP colonies, and previous TS Purple colonies. Digests were performed of P-lambda-R--LacI--TS Purple--no degradation tag/DAS/LAA, R0011--ClpXP, and CI. After analyzing the gel, we concluded the TS Purple with no degradation tag/DAS/LAA, R0011--ClpXP, and CI were of expected sizes, but the P-lambda-R--LacI and R0011--ClpXP--CI digests were unsuccessful. Lastly, the following backbones were successfully digested for future ligations: 1A3, 1C3, 1K3, and 1T3.