October 3-7, 2016

Monday - Friday

Germantown, MD

The Bioscience Education Center

9:00am-5:00pm

1 Hour Lunch Break

This comprehensive lecture/laboratory course is ideal for those bench and research scientists who have a fundamental background in PCR and are looking to gain a thorough understanding of and hands-on exposure to the Real Time, Quantitative and Digital PCR processes.

  • Lecture and Hands-on Interactive Training

  • Team taught by active researchers

  • Comprehensive binder containing workshop material

  • Space limited to 24 participants

  • Registration Fee: $1,095

Course Director

David Kulesh, Ph.D.

Research Microbiologist, U.S. Army Medical Research Institute of Infectious Diseases (USAMRIID)

Lecture Topics:

Intro to real-time PCR; Short History of real-time PCR; Sample Processing (DNA/RNA Extraction and Purification for Real-time PCR); Probe Chemistries and Real-time PCR Instrumentation; Assay Development; Design, function and use of TaqMan & TaqMan-MGB probes; Design, function and use of SimpleProbes and HybProbes; Target selection for real-time PCR; Optimization of TaqMan, TaqMan-MGB, HybProbe and SimpleProbes; Advanced testing (validation) of optimized assays; Medical applications of real-time PCR; Assessing, monitoring, and testing PCR inhibition; Contamination; Quality control; Array qPCR; Multiplex Real-Time PCR; Quantitative Real-Time PCR; Digitial PCR (dPCR);  

Laboratory Topics:

Sample Processing Lab: DNA extraction by manual and automated meth- ods; RNA extraction and Sybr Green Real-Time RT-PCR; 3.      Real-time PCR lab (set up and run DNA and real-time PCR assays on the LC 480 w/ LC 480 Control Kits & Extracted DNA); Analysis of RT-PCR Data; Use of Real-time + PCR Assay Software; Array qPCR (RT and Array Set-up); Analysis of Array qPCR Data; Quantitative Real-Time PCR Lab Set-up; Assemble dPCR Reactions and Initiate Run

Lectures:

  1. Intro to real-time PCR; Short History of real-time PCR
  2. Sample Processing

Laboratory:

  • Sample Processing Lab: DNA extraction by manual and automated meth- ods; RNA extraction and Sybr Green Real-Time RT-PCR

Lectures:

  1. Probe Chemistries and Real-time PCR Instrumentation
  2. Assay Development (Part I); Design, function and use of TaqMan & TaqMan-MGB probes; Design, function and use of SimpleProbes and HybProbes; Target selection for real-time PCR

Laboratory:

  • Real-time PCR lab (set up and run DNA and real-time PCR assays on the LC 480 w/ LC 480 Control Kits & Extracted DNA)
  • Analysis of RT-PCR Data
  • Use of Real-time + PCR Assay Software

Lectures:

  1. Assay Development (Part 2); Optimization of TaqMan, TaqMan-MGB, HybProbe and SimpleProbes; Advanced testing (validation) of optimized assays; Medical applications of real-time PCR; Assessing, monitoring, and testing PCR inhibition; Contamination; Quality contro
  2. Array qPCR
  3. Multiplex Real-Time PCR

Laboratory:

  • Array qPCR (RT and Array Set-up) – Runs overnight

Lectures:

  1. Quantitative Real-Time PCR

Laboratory:

  • Analysis of Array qPCR Data
  • Quantitative Real-Time PCR Lab Set-up
  • Discussion of Quantitative Real-Time PCR Results

Lectures:

  1. Digitial PCR (dPCR)
  2. Wrap-up Lecture

Laboratory:

  • Assemble dPCR Reactions and Initiate Run
  • d-PCR Data Analysis

Dr. David A. Kulesh is a research microbiologist who has been working in the Molecular Diagnostics Department at US Army Medical Research Institute of Infectious Diseases (USAMRIID) for over 16 years. Dr. Kulesh has a PhD and Masters in Biology from The Catholic Uniersity of America (CUA) and a BS from Villanova University. Prior to his employment as a US Army civilian, Dr. Kulesh was a LtCol in the US Air Force stationed at both The Armed Forces Institute of Pathology (AFIP) and USAMRIID. While at AFIP, Dr Kulesh was invloved with studying the effects of microgravity on the differentiation of muscle cells by sending tissue cultures aboard 5 Space Shuttle missions. At USAMRIID, Dr. Kulesh has been responsible for the development of multiple real-time qPCR assays used in the detection of both biothreat bacteria and viruses ranging from anthrax to smallpox to Ebola. His Ebola real-time qPCR assays were used by the US Army during the 2015 West Africa outbreak. Dr. Kulesh has been invlolved in the molecular workshops since he was a graduate student at CUA over 30 years ago.

The Bioscience Education Center

Montgomery College
20200 Observation Drive
Germantown, MD 20876