School of Biotechnology · Molecular biology
Real-Time PCR (qPCR) & Gene Expression Analysis
Quantitative PCR is the standard method for measuring gene expression and nucleic-acid quantity. This course covers the full workflow at the bench and at the computer, so that your numbers stand up to review.
Course overview
Real-Time PCR & Gene Expression Analysis takes you from RNA through reverse transcription to quantitative PCR with SYBR Green chemistry, including primer validation, standard curves, efficiency calculation and melt-curve analysis.
The analysis sessions cover reference-gene selection, the ΔΔCt method and efficiency-corrected models, statistics on biological replicates and the MIQE guidelines that journals expect.
Learning outcomes
By the end of the course you will be able to:
- Prepare cDNA from RNA with appropriate controls
- Validate primers with standard curves, efficiency and melt curves
- Run SYBR Green qPCR plates with replicate and control design
- Calculate relative expression with ΔΔCt and efficiency-corrected methods
- Report qPCR experiments according to MIQE
Skills you will practise
Eligibility and prerequisites
Who can apply
- Students and graduates of B.Sc., M.Sc., B.Tech., M.Tech., B.Pharm., M.Pharm. or a PhD programme in biotechnology, microbiology, biochemistry, molecular biology, genetics, life sciences, pharmacy or allied health sciences
- Laboratory staff, research assistants and faculty who want structured bench training
- Participants must be 18 or older and follow the laboratory safety induction on day one
Prerequisites
- Molecular Biology Techniques or equivalent experience with PCR
Best suited to
- Postgraduate students and researchers measuring gene expression
- Laboratory staff running qPCR assays
- Graduates of the Molecular Biology Techniques course
Module-wise syllabus
5 modules, each with a bench practical performed by every participant.
Topics
- RNA quality and DNase treatment
- Reverse-transcription chemistry
- No-RT and no-template controls
Topics
- Primer design for qPCR
- Standard curves and efficiency
- Melt curves and specificity
Topics
- Plate layout and replicates
- Reference genes
- Instrument settings
Topics
- Baselines and thresholds
- ΔΔCt and the Pfaffl method
- Statistics on biological replicates
Topics
- The MIQE checklist
- Common errors
- Figures for publication
Practical sessions and projects
Practical assessment
Practical assessment
Analyse a qPCR experiment end to end and present fold changes with statistics and a MIQE checklist.
Assessed on technique, accuracy, safety and the laboratory record. A pass is required for the certificate.
- Day 1: RNA to cDNA
Synthesise cDNA from treated and control RNA samples with the full set of controls.
- Day 2: assay validation
Run a dilution series, calculate efficiency and check specificity by melt curve.
- Day 2: the experiment
Run a gene-expression plate comparing treated and control samples.
- Day 3: analysis
Analyse your own plate in a spreadsheet and in R and calculate fold change with confidence intervals.
- Day 3: reporting
Write a MIQE-compliant methods section and figure for your experiment.
Instruments, techniques and materials
Instruments and equipment
Techniques you will perform
- DNase treatment
- Reverse transcription
- SYBR Green qPCR
- Standard curves
- Melt-curve analysis
- Relative quantification
Materials
- Cultured-cell RNA
- Reverse-transcription and SYBR Green kits
- Validated primer sets
Core reading
- Bustin SA et al. (2009). The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clinical Chemistry 55:611–622.
- Livak KJ, Schmittgen TD (2001). Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods 25:402–408.
Duration, format and fees
- Next cohort
- Dates are sent with your admission offer
- Timetable
- Weekdays, 9:30 to 16:30, at our laboratory
- Payment
- Secure online payment link with your admission offer (UPI, cards, net banking). Instalments on request for fees above ₹15,000.
- Refunds
- See the refund and cancellation policy.
Instructor
The instructor for each cohort is named in your admission offer and on this page when the cohort is announced. Vedicgen instructors are selected for hands-on laboratory experience in the subject they teach, and every course is reviewed by a second subject expert.
Questions about who teaches a cohort? [email protected]
Certificate
Learners who complete the course and pass its practical assessment receive a Vedicgen certificate of completion for Real-Time PCR (qPCR) & Gene Expression Analysis, issued by Vivofold, stating the bench hours and techniques performed. Each certificate has a unique ID and QR code that anyone can check.
Apply for Real-Time PCR (qPCR) & Gene Expression Analysis
Apply
Send the form below. It takes about five minutes.
Review
An adviser confirms your fit and format within two working days.
Offer
You receive dates, timetable and a secure payment link.
Onboard
Safety briefing, lab address and joining instructions.
Frequently asked questions
Yes. Day one starts with a safety induction covering personal protective equipment, safe handling of reagents and biological material, spill response and waste segregation. All practicals use non-pathogenic material (Risk Group 1 organisms, established cell lines or synthetic templates) and are supervised throughout.
All reagents, kits, consumables, personal protective equipment, the lab manual and the certificate. Travel, accommodation and meals are not included. The laboratory address and joining instructions are sent with your admission letter.
Bench courses run in small groups so that every participant performs every practical personally, with an instructor-to-participant ratio that allows close supervision at the bench.
Yes. Learners who complete the course and pass its assessment receive a Vedicgen certificate of completion, issued by Vivofold. Every certificate carries a unique ID and a QR code that anyone, including an employer or admissions office, can check on the certificate verification page.
Apply with the form on this page. Live and in-person cohorts run through the year; the dates, timetable and joining details of the next cohort are sent with your admission offer and listed on this page once announced. Self-paced enrolment opens as soon as your application is accepted.
Yes. No course runs longer than four weeks, and the length is customisable: choose the standard schedule, an intensive one- to two-week schedule, weekends only, or a custom timetable. Pick your preference on the application form; colleges and teams can book their own dates.
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