BTEC 2192L Syllabus

Subject Code

BTEC

Course Number

2192L

Course Title

Applied Biotechnology Methods Lab

Prerequisites

BTEC 2191 with a grade of C or higher, BTEC 2191L with a grade of C or higher, CHEM 1211 with a grade of C or higher, CHEM 1211L with a grade of C or higher

Corequisites

BTEC 2192

Terms Offered

Credit Hours

Course Description

This lab course introduces the basic experimental concepts of biotechnology and its associated fundamental laboratory skills. Laboratory activities provide hands-on training in three fundamental areas of modern biotechnology: media preparation and culture of bacteria, isolation and characterization of proteins, and preparation and analysis of recombinant plasmid DNA.

Course Outcomes

Performing Micropipetting and Scientific Calculations As Applied To Chemical and Gel Preparations

  • Demonstrate ability to conduct and practice various micropipetting exercises.
  • Demonstrate proficiency calculating practice scientific math problems as required to prepare various buffers, solutions, media and gels.
  • Develop and maintain a lab notebook detailing protocols, all results, and observations.

Preparing Microbial Media, Solutions and Buffers Using Applicable Laboratory Equipment

  • Prepare complex and rich liquid and solid media.
  • Prepare selective and differential media.
  • Prepare minimal media.
  • Prepare antibiotic solutions and media additives.
  • Prepare biological buffers using a pH meter.
  • Demonstrate sterile techniques when preparing media using autoclaves and sterile filtration techniques.

Isolating Specific Organelles and Conducting Marker Enzyme Assays

  • Prepare a homogenate suitable for differential centrifugation to isolate organelles.
  • Determine the total protein concentration of your organelle prep using a Bradford assay.
  • Examine and assay the organelle prep for malate dehydrogenase activity.
  • Calculate the rate of malate dehydrogenase followed by determining its specific activity.
  • Create Excel graphs to illustrate Bradford results and various assay slopes.

Partially Purifying and Characterizing Proteins Using Columns, Assays and Protein Gel Analysis

  • Demonstrate the ability to pack and equilibrate sephadex columns.
  • Demonstrate the ability to run sephadex columns to purify a bioluminescent protein.
  • Demonstrate the ability to denature the purified protein and investigate bioluminescent activity.
  • Examine and analyze native and denatured protein preps using PAGE determining the molecular weights of unknowns.
  • Prepare and set up a functional polyacrylamide gel system.
  • Explain how to visualize and analyze proteins in a polyacrylamide gel.

Isolating Plasmid DNA  and Gel Analysis

  • Demonstrate the ability to grow bacteria harboring plasmid DNA in selective medium to saturation.
  • Demonstrate the ability to isolate plasmid DNA from saturated culture.
  • Prepare an agarose gel using prepared agarose stock and electrophoresis buffer.
  • Demonstrate the ability to run and analyze plasmid DNA using prepared agarose gel.
  • Demonstrate the ability to visualize and photo-document DNAs in gel using stain and ultraviolet light.
  • Explain the results of agarose gel electrophoresis using known control DNAs.

Restricting DNA and Analysis; Ligation and Transformation of Bacteria Using Selection

  • Perform a restriction analysis on the isolated plasmid DNA.
  • Demonstrate the ability to analyze restricted plasmid DNA using proper size controls and agarose gel analysis.
  • Perform a ligation of restricted plasmid DNA.
  • Perform chemical transformation using a proper bacterial host strain, ligation mix and appropriate selection.
  • Calculate transformation results in context with proper transformation controls.