Blog

Blog

Guides, tutorials, and best practices for qPCR data analysis.

What Happened to qbase+ — and Why I Built VoilaPCR

I went looking for qbase+ and it was gone. So I built a browser-based qPCR analysis tool that does what qbase+ did, without the pain.

qPCR Standard Curve Best Practices and Common Mistakes

How to build a reliable qPCR standard curve: dilution series setup, efficiency calculations, and the mistakes that quietly wreck your absolute quantification.

How to Normalize qPCR Data with Multiple Reference Genes

Using multiple reference genes in qPCR improves normalization accuracy. Here's how to combine them correctly using geNorm, NormFinder, or a simple geometric mean.

Multiplex qPCR: A Practical Guide to Running Multiple Targets in One Well

How to design, optimize, and troubleshoot multiplex qPCR — primer/probe design, spectral crosstalk, efficiency matching, and common pitfalls.

How to Design qPCR Primers That Actually Work

Practical primer design rules for qPCR: Tm matching, amplicon length, specificity checks, and the validation steps most protocols skip.

The Pfaffl Method: When Your Primer Efficiencies Make ΔΔCt Unreliable

When qPCR primer efficiencies differ between target and reference genes, the Pfaffl method gives you accurate fold changes. Here's how to use it.

Melt Curve Analysis in qPCR: What It Tells You and When to Worry

How to read melt curves in SYBR Green qPCR, spot primer dimers, identify off-target products, and decide when a run is trustworthy.

SYBR Green vs TaqMan: When to Use Each Chemistry

A practical guide to choosing between SYBR Green and TaqMan for your qPCR experiments — based on cost, specificity, and what your project actually needs.

Choosing the Best Reference Genes for qPCR

How to pick, validate, and use the right housekeeping genes for accurate normalization in RT-qPCR experiments.

How to Calculate DDCt (Livak Method): A Complete Guide

Learn the delta-delta Ct method step by step, including the math, assumptions, common mistakes, and worked examples.

qPCR Troubleshooting Guide: 10 Common Problems and Fixes

A practical checklist for diagnosing and fixing the most common qPCR issues, from NTC amplification to unexpected fold changes.