PCR

Find guides, protocols and helpful tips when performing the polymerase chain reaction (PCR).

Pfaffl results graph

How To Perform The Pfaffl Method For qPCR

What is the Pfaffl method? The Pfaffl method, named after it's curator Michael Pfaffl, is used to calculate relative gene expression data while accounting for...

13 Free PCR Primer Design Programs

Gone are the days where you have to design primers by hand. Instead, we have a plethora of excellent resources available that utilises complex...

How To Make TE Buffer pH 8.0

About TE buffer TE buffer (Tris-EDTA) is a commonly used buffer solution for resuspending and storing nucleic acids, especially DNA. The Tris solution keeps the...

How To Dilute New PCR Primers

In this article, I will explain how to easily prepare PCR stock primers and how to dilute them into a working primer solution. So you...

How To Make TBE Buffer

About TBE buffer TBE buffer, named so because of the three ingredients of Tris base, Boric acid and EDTA, is a solution commonly used as...
Bisulfite pyrosequencing pyrogram

How Does Bisulfite Pyrosequencing Work?

What is bisulfite pyrosequencing? Bisulfite, also referred to as bisulphite, pyrosequencing is a technique used to quantify DNA methylation levels on DNA at a single-base...
qPCR amplification plot annotated

What Is A Cycle Threshold (Ct) Value In qPCR?

In this article, I will explain what is meant by a Ct value in quantification (real-time) PCR (qPCR). I will also discuss the difference...

What Is The Polymerase Chain Reaction (PCR)?

What is the Polymerase Chain Reaction (PCR)? PCR is a molecular technique frequently applied to amplify regions of interest in DNA, resulting in millions or...
Random primers, oligo(dT) and gene-specific primers for cDNA synthesis

Random Primers Or Oligo(dT) Primers For cDNA Synthesis?

Having extracted and assessed the total RNA from your samples, you now have the task of creating the first strand complementary DNA (cDNA). Sounds...
Agarose gel

Reasons Why Your PCR Reaction Does Not Work

We have all been there. You have been performing PCRs perfectly for weeks/months on end but now, for some bizarre reason, you see …...