PCR

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

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 Make TBE Buffer (10x)

About TBE buffer TBE buffer, named so because of the of the three ingredients of Tris base, Boric acid and EDTA, is a solution commonly...
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...
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...
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...
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...
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How To Perform The Delta-Delta Ct Method

What is the delta-delta Ct method? The delta-delta Ct method, also known as the 2-∆∆Ct method, is a simple formula used in order to calculate...

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...
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 …...
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How To Create Real-Time PCR Primers Using Primer-BLAST

Primer-BLAST Primer-BLAST is NCBI’s free online primer design platform which designs PCR primers using the Primer3 system and simultaneously checks the likelihood that the primers...

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