National Heart, Lung, and Blood Institute program in
Acute Lung Injury
Specialized Centers of Clinically Oriented Research (ALI SCCOR)

University of Washington ALI SCCOR Program
Translational Research Training Program

Laboratory Skills Workshop

Course Director: Lynn M. Schnapp, MD

Day 1
Lecture: Intro to Lab Etiquette (how not to destroy the lab) Introduction to Protein Work
Group A	Group B
Make cell lysates	Isolate plasmid DNA
Measure protein concentrations	Measure DNA concentration
Set up protein gel	Run on agarose gel
Load samples	Perform Restriction Digest
Run samples	Analyze by agarose gel
Transfer	(Make cell lysates)
Block membrane
(Pick colonies into liquid culture)

Day 2
Lecture: Intro to DNA/Plasmids/RE
Group A	Group B
Primary antibody incubation	(Make cell lysates)
Secondary antibody incubation	Measure protein concentrations
ECL	Set up protein gel
During incubations	Load samples
Isolate plasmid DNA	Run samples
	Transfer overnight

Day 3
Lecture: Antibodies
Group A	Group B
Measure DNA concentration	Block membrane
Prepare agarose gel	Primary antibody incubation
Run plasmid DNA on agarose gel	Secondary antibody incubation
Perform Restriction Digest	ECL
Analyze by agarose gel

Day 4
Lecture: PCR
Group A	Group B
Reverse Transcription	ELISA
Set up PCR reactions	Reverse Transcription
Analyze PCR reactions on gel

Day 5
Lecture: Going in Other Directions: Southerns, Northerns, etc.
Group A	Group B
ELISA	Set up PCR reactions
	Analyze PCR reactions on gel

Appendix: Practical Solution Making

Hit Counter

For problems or questions regarding this Web site contact webmaster@alisccor.org
Last updated: 12/16/05.