MCCCD Official Course Competencies:

BIO212AA  20026-20045

Biotechnology I

1.

Define the term "biotechnology," and trace its history as a field of science. (I)

2.

Describe the role of biochemistry and genetics in cell function and biotechnology. (I)

3.

Explain the concepts of molarity, normality, and % by performing basic mathematical calculations to prepare solutions and media. (II)

4.

Define the concept of pH, and explain how a pH meter functions. (II)

5.

Select appropriate media type for various cell types. (II)

6.

Perform spectrophotometric analysis by determining concentration of unknown DNA samples. (III)

7.

Monitor microbial growth by generating a growth curve from a growing culture. (III)

8.

Describe industry established standards for record-keeping in the laboratory, and perform daily record-keeping. (IV)

9.

Isolate genomic DNA and purify plasmid DNA using current purification protocols. (IV)

10.

Explain the concepts of transformation, transfection, transduction, plasmids, phagemids, and clones. (V)

11.

Examine samples of purified DNA by agarose gel electrophoresis and spectrophotometric analysis. (V)

12.

Explain recombinant DNA experiments that led to the advent of biotechnology as a scientific discipline. (VI)

13.

Explain promoter and regulator function in gene expression by designing a novel expression vector. (VII)

14.

Present evidence of successful ligation between fragmented genomic DNA and vector DNA to create a plasmid library. (VII)

15.

Generate a restriction map for the recombinant selected from a plasmid library. (VIII)

16.

Design primers to amplify cloned fragment from genomic DNA. (VIII)

17.

Explain southern, northern, and western blot hybridization. (VIII)

18.

Describe at least three different methods of DNA sequencing. (VIII)

19.

Apply knowledge of promoters in the design of a hypothetical promoter-probe vector. (IX)

20.

Prepare DNA for sequencing by sub-cloning fragment selected from plasmid library in pUC19 and purifying ssDNA using M13 helper phage. (X)

21.

Provide evidence of successful sub-cloning by southern blot hybridization. (X)

MCCCD Official Course Outline:

BIO212AA  20026-20045

Biotechnology I

I. Modern Biotechnology

A. What is it?

B. Ancient biotechnology

C. Classical biotechnology

D. Foundations of modern biotechnology

E. Industry standards for record-keeping

II. Media and Solution Preparation

A. Units of concentration and volume

B. Basic mathematical calculations for laboratory science

C. Buffers and pH

D. Autoclaving and filter sterilization

E. Growth media for prokaryotic cells and for eukaryotic cells

F. Differential and selective media

III. Basic Principles of Recombinant DNA Technology

A. Digesting and ligating DNA

B. Separating restriction fragments and visualizing DNA

C. Cloning vectors

D. UV spectroscopy: determining DNA concentration and purity

E. Microbiology's role in biotechnology

F. Growth curve

G. Phases of growth

IV. Genomic DNA

A. Physical characteristics of genomic DNA

B. Isolating genomic DNA from E. coli

V. Competent E. coli

A. Transformation, transfection, and transduction

B. Preparing E. coli to accept foreign DNA

C. Plasmids, phagemids, plaques, and clones

D. Vector

E. Transformation efficiency calculations

F. Purification of plasmid DNA

VI. The DNA Revolution

A. The first recombinant DNA experiments

B. Safety concerns

C. Current and future concerns

VII. DNA Library: A Collection of Life's Secrets

A. Constructing and screening a DNA library

B. Screening libraries and expression libraries

C. Genomic libraries and cDNA libraries

D. Regulation of gene expression

E. Understanding and exploiting gene expression

F. Promoters and regulators

G. Restriction digestion and multiple cloning sites

H. Recombinants

VIII. Characterization of Recombinants

A. Restriction mapping

B. PCR

C. Southern, northern, and western blot hybridizations

D. DNA sequencing

IX. Promoter Strength

A. Reporter genes

B. Characteristics of reporters

C. Enzyme assays to compare strength of different promoters

D. ? galactosidase and lacZ

X. The DNA Sequencing Template

A. Helper phage

B. Sub-cloning

C. Infections

D. Single-strand DNA purification

E. Southern blot hybridization