BISC 431 – Molecular Biotechnology

Course Outline – (2002)

Instructor: Dr. Robert W. Sharp – “Rob Sharp”

Office: B9232

E-mail: rwsharp@sfu.ca

Phone: 604 291 5626

Lectures: Tuesday 10:30-11:20, Room C9000

Thursday 9:30-11:20, Room C9000

Labs: Tuesday, Wednesday, Thursday, 13:30-17:20, SSB 8133

Office Hours: Tuesday 11:30-12:20, Wednesday 10:00-11:00, Thursday 11:30-12:30

Description: Laboratory/Lecture molecular biotechnology course

Textbook: Howe, C. Gene Cloning and Manipulation

additional materials will be placed on reserve in the library

Lab experiments:

Plasmid and genomic DNA preps, recombinant DNA techniques, Southern blots, PCR, sequencing, and sequence analysis will be performed in lab exercises. Some labs may extend for more than one lab period, and may require participation outside of the scheduled lab period. Study questions will be included in lab handouts – answers to these will be due during the next lab period. A full lab report on the term’s experiments will be due in the final week of classes. Students will also be expected to keep an up-to-date, neat, lab book (dated and written in pen). Lab books will be requested at random intervals, without prior warning.

Lab Schedule (Titles of labs will be converted to hyperlinks to lab handouts)

Week	Dates	Lab
1	Tues. Jan. 15-Thurs. Jan. 17	Phage dilutions
2	Tues. Jan. 22-Thurs. Jan. 24	Genomic DNA prep
3	Tues. Jan. 29-Thurs. Jan. 31	Plasmid miniprep sample gel
4	Tues. Feb. 5- Thurs. Feb. 7	Southern blot Lab 4/5 and 7 results
5	Tues. Feb. 12-Thurs. Feb. 14	set up PCR reactions
6	Tues. Feb. 19-Wed. Feb. 20	no labs – reading break
7	Tues. Feb. 26-Thurs. Feb. 28	gel of PCR products
8	Tues. Mar. 5- Thurs. Mar. 7	Subcloning into plasmid vector results
9	Tues. Mar. 12-Thurs. Mar. 14	DNA sequencing
10	Tues. Mar. 19-Thurs. Mar. 21	edit sequence
11	Tues. Mar. 26-Thurs. Mar. 28	computer analysis of sequence
12	Tues. Apr. 2- Thurs. Apr. 4	Full lab report due Lab report format

Lab Safety Information General Safety Tips

Aseptic technique and equipment use Chemical and radiation safety

Lecture topics:

Lecture topics will mostly be related to laboratory material. Topics covered will include recombinant DNA techniques, PCR, gene expression, and mutagenesis. A library research project will also be due in the first week of March.

Lecture notes: (Click on links below for lecture notes)

(These are outline notes only – supplement with notes in lectures and text readings)

Introduction/overview

Preparing DNA for cloning – overview

vectors – plasmids

M13 phage derivatives

λ vectors – introduction

– cloning vectors

– cloning in lambda

– in vitro packaging

vectors for cloning large DNA fragments

– preparation of large DNA fragments

preparation of foreign DNA – restriction enzymes

sample mapping question

preparation of probes – oligonucleotide synthesis

cDNA

RACE

screening – Southern blots

– probes

cloning – ligation and transformation

PCR – special applications

gene analysis and mapping – restriction mapping

sequencing longer sequences genomic sequencing strategies

gene identification genomics

Grading:

	Date	Time	Percent of final grade
Midterm	February 14	10:30	15 %
Final	Friday, April 12	8:30-11:30	50 %
Lab study questions	various	17:20	8 %
Lab note book	various	13:30	2 %
Full lab report	April 2-5	17:20	15 %
Library research project	~ March 5	10:30	10 %

Lab Safety Information General Safety Tips

Time