Biol Chem 243 Overview

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Biol/Chem 243 - Nucleic Acids Lab

Biol/Chem 243 - Overview

Spring 1999

Biology/Chemistry 243 is a 3-unit graduate level course. The prerequisites are Biol/Chem 241AB, or current registration in 241B. This course meets on Tuesday from 1 to 5 and on Thursday from 2- 5. The actual hours for the lecture portion of the class will be flexible within the hours listed above. Due to the nature of the molecular biology laboratory, students must be able to put in occasional hours outside of normally scheduled lab times in order to complete some of the procedures.

Instructor: Alice D. Wright

10B Science 278-7692

Office hours: Tuesday and Friday 10:30 am to noon

Email: awright@csufresno.edu

Texts:

Gene Cloning, 3rd edition. 1995. T. A. Brown. Chapman and Hall, publishers. DNA: A project Approach. 1995. Karcher. Academic Press, publisher.

Goals:

The objectives of this class are to provide students with hands-on experience with current molecular biology techniques and the science behind those techniques and to make the student feel comfortable with incorporating those techniques into his or her own research project, should that be appropriate.

Grading:

30% lab reports (2 will be turned in 15% each)

1. cloning 1-28 to 2-23 due 3-5

2. your choice

a. shot gun cloning 2-25 to 3-18

b. Northern blot 4-15 to 5-4

25% class participation / lab notebook (student provides lab notebook)

25% final examination (take home test)

20% student presentations 30 to 45 minutes

Covering a procedure that we did not do in class, using at least one example of the procedure in current literature (with in the last year).

If you have special needs as addressed by the Americans with Disabilities Act and need course materials in alternative formats, notify your instructor immediately. Reasonable efforts will be made to accommodate special needs.

Biol/Chem 243 - Lab Syllabus

Spring 1999

LAB SCHEDULE:

Some modifications of procedures will be made to fit the experiment. Students should come to lab prepared to perform the experiment - having read and understood the directions and having prepared the necessary solutions. There will be time in previous lab classes to prepare the solutions for the next lab.

Date Lecture reading from Brown LAB Reading/procedure( Karcher)

1-26

Introduction, safety, plasmids
2.1, 3.2, 6.1

make solutions, buffers

read chapter 2

1-28 plasmid isolation 2.6a &b

2-2 Enzymes, chapter 4 gel electrophoresis 2.2

2-4 restriction enzyme digestion, gel electrophoresis 2.1a

2-9 transformation, 5.1, 5.2 isolation of DNA from gel, ligation 2.4

2-11 transformation of E. coli 2.4

2-16 DNA isolation, Chapter 3 terile techniques, isolation p. 12-15

2-18 plasmid preps

2-23 cloning vectors, chapter 6 Restriction endonuclease digestion, gel

2-25 chromosomal DNA preparation instructor supply

3-2 cloning vectors, chapter 7 partial Sau3A digestions, gel read chapter 3

3-4 transfection of E. coli, make DNA probe adapt from 3.3

3-9 selection, chapter 8 replica plating, selection p. 27-29

3-11 colony hybridization 3.1d

3-16 gene and genome structure,
Chapter 9 hybridization and washing next day 3.4d

3-18 detection

3-23 structure continued PCR read chapter 6, 6.1

3-25 presentation topics due analysis of PCR products

3-29 TO 4-2 SPRING BREAK

4-6 mobile genetic elements Transposon mutagenesis read chapter 1

4-8 isolation of auxotrophs 1.5

4-13 gene expression, chapter 10 identification of auxotrophs 1.6

4-15 RNA extraction read chapter 5, 5.1

4-20 gene expression continued RNA extraction, separate poly-A RNA 5.2

4-22 run gel and set up blot 5.3, 5.4

4-27 TBA make probe for Northern 5.6

4-29 hybridization (washes next day)

5-4 sequencing detection of probe

5-6 DNA sequence analysis

5-11 student presentations

5-12 student presentations