Week 11 — Building Genomes
From DNA design to building genomes and bioproduction: this week bridges sequence design with practical pathways to construct systems that produce carotenoids and other compounds.
- Info Slides will be posted here after class; recording will appear once processed. Recording: Zoom
- Warning Homework will be posted here when available. Use this week to deepen your strain design plans and connect them to next week’s imaging/measurement workflows.
- Info Recitation slides will be shared here after the class; recording will be linked below. Recording: Zoom
- Tip In this two‑day lab, you’ll produce beta‑carotene and lycopene in E. coli and analyze growth (OD600) and pigment absorbance. Objective Work with E. coli carrying pAC‑LYC (lycopene) and pAC‑BETA (beta‑carotene) to optimize pigment production by tuning media, temperature, and carbon source. Concepts You’ll Learn Fundamentals of bioproduction Tuning environmental factors to optimize production Measuring OD600 and absorbance spectra Handling cultures, plasmid systems, and acetone extraction Key Plasmids pAC‑LYC (Addgene #53270) pAC‑BETA (Addgene #53272) Workflow (2 Days) Day 1: Set up overnight cultures; vary LB vs 2YT, 37 °C vs 30 °C, and ± fructose. Day 2: Measure OD600; extract pigments with acetone; record absorbance spectra for lycopene vs β‑carotene.
- pAC‑LYC — Addgene pAC‑BETA — Addgene Bioproduction with fructose — Biotechnol Lett (2016) OD600 & absorbance basics — ThermoFisher protocol