qPCR course information

Quantifying nucleic acids by real-time PCR – Practical Workshop


Next workshop: Thursday and Friday 23rd & 24th November 2017

£460 academic participants, £560 other participants

Earlybird registration deadline 27th October 2017. After this deadline an additional £100 fee applies.
Scroll down to see a summary of the feedback from the previous two years’ participants and workshop timetable.

Register here for this hands-on course  (you will be asked to confirm that you have read the Terms and Conditions before registering).This two-day workshop is aimed at those who would like to use real-time PCR to quantify DNA or mRNA. It has evolved from workshops we have run over the past 14 years for the BBSRC Molecular Training for Industry programme, King’s College graduate school and The Physiological Society. Participants are expected to have a theoretical understanding of the polymerase chain reaction and reverse transcription. No prior knowledge of real-time PCR is expected. Accommodation and travel costs are not included in the price. This workshop will be held at University College London, Gower Street.  For more information on payment methods and how to get there please visit the links to the right.

Feedback summary for 2014 & 2015 (n=106)

What was your overall evaluation of the workshop?
Poor (0) 0%
OK (3) 2.8%
Good (32) 30.2%
Excellent (71) 67.0%

Did you find that 2 days was –
Not enough time (20) 18.9%
Too long (2) 1.9%
About the right length (84) 79.2%

The practical and theoretical material covered in the workshop –
Was completely new to you (30) 28.3%
Complemented previous experience (28) 26.4%
Added to previous experience (47) 44.3%
Repeated previous experience (1) 0.9%

Did you attend this workshop because –
Of general interest in qPCR (9) 8.4%
You may use qPCR in the future (21) 19.8%
You will be using qPCR soon (40) 37.7%
You are already using qPCR (36) 34.0%

Payment methods

How to get there (UCL)

How to get there (Edgware)


Local places of interest




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Course content

Workshop Day 1 – 09.45-17.30

Sample/template preparation

Theory: Isolation of DNA, total RNA and mRNA. Assessing template quantity and quality prior to qPCR.

Practical: Isolation of total RNA from tissue using spin columns. Analysis of total RNA quantity using a Qubit fluorimeter, and RNA quality using an Agilent Bioanalyzer.

Reverse transcription (RT) of mRNA and preparation of known copy number standards

Theory: Use of total RNA or mRNA. RT priming strategies and enzymes.

Practical: Set up RT reaction with total RNA isolated earlier. Set up a qPCR assay with the newly-synthesised cDNA. Check PCR product specificity/identity by agarose gel electrophoresis.

Assay design

Theory: qPCR assay development; primer design; product detection strategies; PCR conditions/optimisation.

Standard operating procedures

Theory: How to avoid contamination

Workshop Day 2 – 09.15-16.30


Practical: Set up and run a quantititive probe assay. Prepare a standard dilution series. Set up and run a quantitative SYBR Green assay with standards, cDNA samples, and positive and negative controls.

Data analysis

Practical: Evaluate assay efficiency, sensitivity, reliability, accuracy. Compare results from probe and SYBR assays.

Theory: Aspects of data analysis: Normalisation of qPCR data – demonstration of geNorm, absolute versus relative quantification, quality control and MIQE.

Practical: Analyse a provided RT-qPCR dataset. Evaluate assay efficiency, select reference genes for normalisation, normalise data and compare group means (a limited number of computers will be provided but we ask participants to bring a PC laptop if possible).

Theory: Troubleshooting and questions.