Hands-on Real-Time PCR Workshop: A Complete Course for Successful Gene Quantification & Genotyping
Gene Expression Analysis Series
Date: April 6-7, 2005
Time: See the schedule
Location: Henry Koffler Building 510
Instructors: Paul Mola and Jennifer Biesterfeldt
Cost: $50
Seating: Limited
Registration: Required
Flyer: Open the real-time PCR flyer
REGISTRATION CLOSED
Description:
This is an intermediate to advanced course for scientists familiar with PCR. Real-time reverse-transcriptase (RT) PCR quantitates the initial amount of the template most specifically, sensitively and reproducibly, and is a preferable alternative to other forms of quantitative RT-PCR that detect the amount of final amplified product at the end-point. The technology is especially useful for evaluating "RNA phenotypes" obtained from microarray, siRNA or gene expression experiments. The course will cover absolute and relative real time qPCR strategies including hands on experimentation. We will address quality control issues including RNA preparation and cDNA synthesis.
Gene assay design software will be covered in detail. Also covered are choice, construction and evaluation of qPCR standards, optimization and troubleshooting of quantitative PCR, experimental design and setup. Lectures will include theory and background, advanced concepts and applications.
You will have an opportunity to get advice and design your PCR primers prior to the session. After performing the PCR reactions, we will then evaluate and learn to interpret the results. Troubleshooting the experiment will also be done plans can be made for the next set of experiments.
Requirements:
- Sequence information about the genes to examine for designing the PCR primers
- You will need to purchase the primers prior to the workshop
- PCR primers (if you've already designed them)
- You must bring your own PIPETTORS
- You must bring your own BARRIER TIPS
Schedule:
Wednesday 06Apr2005
9:30-11:30
11:30-1:30 Lunch Break
1:30-4:00Thursday 07Apr2005
9:30-11:30
11:30-1:30 Lunch Break
1:30-4:00
Tentative Agenda:
Day 1
Sample preparation and nucleic acid isolation and extraction
The dos and don'ts of tissue homogenization
Column vs. other methods of nucleic acid extraction
Assay design and development including
Primer design
Choice of chemistry
Assay optimization considerations
Wet lab
Nucleic acid extraction
Reverse transcription
Real-time PCR
Prior to the workshop - primer design and consultation will be offered to provide primer sequences for primers that will be used during the workshop
Day 2
Analysis, interpretation and reporting of real-time gene expression results
Absolute quantification with external standard curves
Relative quantification with a house keeping gene
Relative quantification with a house keeping gene and calibrator
Relative quantification with efficiency correction
Troubleshooting
What next?
This workshop is offered by GATC and 
Need additional info?
Contact Al Agellon agellon@u.arizona.edu
Limited Seating, Registration Required.
