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Real-time PCR Analysis

Introduction

Real-time PCR, a leading tool in many laboratories for nucleic acid amplification and detection, is among the most valuable techniques used in biological research today. Scientists in all areas of research — basic science, biotechnology, forensic science, diagnostics, biomedicine, microbiology, veterinary science, agriculture, pharmacology and toxicology rely on this method. Real-time quantitative PCR allows the sensitive, specific and reproducible quantitation of nucleic acids. Since its introduction, real-time quantitative PCR has revolutionized the field of molecular diagnostics and the technique is being used in a rapidly expanding number of applications.

It offers interesting new applications, such as for the quantification and genotyping of pathogens, gene expression, methylated DNA and microRNA analysis, validation of microarray data, allelic discrimination and genotyping (analysis of SNPs and microsatellites, identification of chromosomal alterations), validation of drug therapy efficacy, forensic studies and quantification of gene expression in genetically modified organisms (GMOs).

Real Time PCR technology merges the polymerase chain reaction chemistry with the use of fluorescent reporter molecules in order to monitor the production of amplification products during each cycle of the PCR reaction. In addition to quantification experiments, qPCR can be used for the analysis of copy number variations, DNA methylation, and genomic variants.

This exciting technology has enabled the shift of molecular diagnostics toward a high-throughput, automated technology with lower turnaround times. Thus Keeping in mind, the requirements of today’s research and industry, we maintain state of the art instrumental setup with a competent team where customized qPCR projects always help students to find new endeavor in global world and stood confidently for preparing for interviews or applying for PhD across the world. There is huge opportunities for well trained qPCR person for gene expression studies in Genomics, Bio-pharmaceuticals, Vaccine production industries, Human and Animal diagnostic centres, Bioprocessing units and many more…

Course Overview:

A combination of lectures and hands on laboratory experience in using real-time PCR methods. Gain experience using DNA binding dyes and hydrolysis probe chemistries as well as real-time PCR. Lectures include an overview of gene expression, assay design and optimization, analysis of results, applications of real-time PCR technology, and more

Course I: Workshop in Real-time PCR

Course code: RTPCR-WS

Duration: 1 day

Batch Timing: 10 am- 6 pm( 2nd and 4th Saturday every month)

Batch size: 10 students

Course Information:
One day training program that provides training about Real-time PCR Basics like Principle, Instrumentation, Applications  demonstration in Lab by experts on sample acquisition and analysis. (Batch of 10 students only and Certificate of Attendance of Real-time PCR workshop)

Course Fees: 1500 /-

Name of the Topic

Content Covered

Introduction to Real time PCR

 Basic Principle of Polymerase Chain Reaction and Real time PCR.

 Working and Principle of Instrument.

Applications of Real time PCR.

Gene expression analysis:

Sample preparation

Analysis

Introduction of multi gene PCR.

Sample preparation and reaction set up for DNA binding dye chemistry

Protocol set up in QS5 and data collection

Result Interpretation Result Analysis and study of gene expression pattern
Recap Session Question and Answer Session and Certificate distribution

Course II: CERTIFICATION COURSE IN REAL TIME PCR

Course Code: RTPCR-ATR

Duration: 4 days

Batch Timings: 4-6pm Tuesday to Friday

Batch Size: 5 students.

New batch starts: Every 2nd and 4th Tuesday every month

Course Information: Real Time PCR Advance Hands-on Training

It is a combination of lectures and hands on laboratory experience in 3 days training session. The course involves Basics of Real time PCR including Principal, Instrumentation, DNA binding dyes and Taq man probe chemistries to study gene expression, Applications and Complete hands on in cDNA preparation, reaction set up with data analysis for DNA binding dyes and Taq man probe chemistries. Certification of completion of Advance Hands on Training in Real time PCR provided at the end of the training.

Course Fees: 4500 /-

Course Overview:

Name of the Topic

Content Covered

DAY 1

Lecture I: Introduction to Quantitative Real-time PCR

  • Basic Principle of Polymerase Chain Reaction and Real time PCR
  • Working and Principle of Instrument
  • Applications of Real time PCR
  • Expt 1: cDNA preparation
DAY 2

Expt 2: Gene Expression analysis of target gene by using SYBr fluorescence chemistry

 

  • Introduction of multi gene PCR and line of reference generation
  • Real-time PCR of target gene and reference gene
DAY 3

Expt 3: Gene Expression analysis of target gene by using Taq-Man Probe Chemistry

  • Introduction of multi gene PCR and line of reference generation
  • Real time PCR of target gene and reference gene
  • Calculations and analysis ofExperiment no.2
DAY 4

Result Interpretation

  • Calculations and analysis of Experiment no.3
  • Vote of thanks and Certificate distribution

Ongoing Projects:

Introduction to high throughput gene expression analysis:

Gene expression analysis is most simply described as the study of the way genes are transcribed to synthesize functional gene products — functional RNA species or protein products. In addition to quantification experiments, qPCR can be used for the analysis of copy number variations, DNA methylation, and genomic variants. Here, we performed an extensive characterization of the workflow by evaluating the Cells-to-Ct value, and benchmarked it to the hither to gold standard workflow of RNA purification and qPCR gene expression studies.