Biotechnology Design I - Labs

Biotechnology Design I Laboratory Modules

Laboratory 1 : Microbiology Basics & Protein Expression 1. 

Key Pre-Lab Videos

1. Pouring Agar Plates (8:49)

2. Isolation Streaking for Single Colonies (1:36)

3. Basics of Protein Expression in E. coli  (8:22)

4. Overview of Proteins we are expressing - Taq (1:23)

5. Overview of Proteins we are expressing - Cas9 (2:09)

6. Composition of E. coli (Biomass) (Biochemical Eng Fund, Lecture 2) (19:54)

Lab Checklist

PreLab Quiz (Full Credit Taken Before Lab, 50% Maximum After; Answer Key)

Lab Protocol 

Lab Data Sheet (None for Lab 1)

Laboratory 2 : Protein Expression 2, Analyzing Expression Levels, Cell Harvest

Key Pre-Lab Videos

1. SDS PAGE (3:16)

2. Image J (3:29)

3. Overview of Fermentation (15:03)

4. Downstream Processing Basics 1 (11:03)

5. Downstream Processing Basics 2 (10:08)

6. Centrifuge Design (20:41)

PreLab Quiz (Full Credit Taken Before Lab, 50% Maximum After; Answer key)

Lab Checklist

Lab Protocol (Example gel picture in lab)

Lab Data Sheet 

Lab Report 1 Guideline & Rubric (Due Sep 20th)

Laboratory 3 : Protein Expression 3, Cell Lysis & Basics of DSP

Key Pre-Lab Videos

1. Readings on sonication and other types of cell lysis methods. (1, 2)

2. Brief instruction of sonication (2:00)

3. Brief intro of BugBuster reagent (1:36)

4. Overview of Analytical Methods (6:41)

5. Overview of Bradford Assay (11:53)

6. Cell Disruption/Homogenization (3:49)

Lab Checklist

Lab Protocol

PreLab Quiz (Full Credit Taken Before Lab, 50% Maximum After; Answer key)

Lab Data Sheet 

Laboratory 4 : Downstream Purification 2: His Tag Purification 

Key Pre-Lab Videos

1. Overview of His-tag Protein Purification (4:51)

2. Overview of Gravity Flow Column Chromatography (5:37)

3. Extra Information on His-tag purification (1, 2; You may need these for the quiz!)

Lab Checklist

Lab Protocol

PreLab Quiz (Full Credit Taken Before Lab, 50% Maximum After; Answer key)

Lab Data Sheet 

Laboratory 5 : Downstream Purification 3: Protein Precipitation 

Key Pre-Lab Videos

1. Overview of Effect of Temperature on Protein Folding 

Lab Checklist

Lab Protocol 

PreLab Quiz (Full Credit Taken Before Lab, 50% Maximum After)

Lab Data Sheet 

Lab Report 2 Guideline & Rubric (Due Oct 18th)

Laboratory 6 : Downstream Purification 4: Tangential Flow Filtration (TFF)

Key Pre-Lab Videos

1. Overview of GFP Purification by TFF #1 (1:22)

    Overview of GFP Purification by TFF #2 (1:47)

    Overview of GFP Purification by TFF #3 (0:58)

    Overview of GFP Purification by TFF #4 (0:58)

    Overview of GFP Purification by TFF #5 (1:42)

    Overview of GFP Purification by TFF #6 (1:51)

For reference:

2. Installation of the TFF system we use in lab

3. Manual of the TFF system we use in lab

PreLab Quiz (Full Credit Taken Before Lab, 50% Maximum; Answer Key)

Lab Protocol 

Laboratory 7 : Downstream Purification 4: Chromatography 

Key Pre-Lab Videos

1. Overview of FPLC

2. Types of Chromatography use for Proteins:

   - Size Exclusion Chromatography (1:04)

   - Ion-exchange Chromatography (1:40)

   - Affinity Chromatography (1:11)

3. Overview of Protein Size and pI

4. Modelling Chromatography 

5. Chromatography Design 

PreLab Quiz (Full Credit Taken Before Lab, 50% Maximum After;)

Lab Protocol 

Lab Report 3 Template

Laboratory 8 : Protein Quality Control & Enzyme Kinetics 

Key Pre-Lab Videos

1. Overview of qPCR (2:44)

2. Overview of cas9 digestion (6:21)

PreLab Quiz (Full Credit Taken Before Lab, 50% Maximum After)

Lab Protocol 

Lab Report 4 Template