Techniques / PCR, qPCR and qRT-PCR

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droplet digital PCR read on

Divide and Conquer: How to Setup Your First Droplet Digital PCR Experiment

Droplet digital PCR?  It’s easy. Because we’re here to guide you through it. We recently introduced you to the principles of digital PCR technology and how it differs from qPCR. In a nutshell, digital PCR is an end-point PCR technology that divides a single PCR into a large number of partitions, and then perform PCR […]

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In PCR, qPCR and qRT-PCR 6th of April, 2017
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RT-qPCR read on

How to Choose the Correct Reverse Transcription Method

Quantitative Reverse transcription PCR (RT-qPCR) is frequently used in the lab to detect and quantify RNA expression in a sample. The first step of the assay is to convert the labile RNA to its complementary DNA (cDNA) counterpart through reverse transcription (RT). In fact, RT is the first step in a variety of molecular biology […]

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Reverse Transcription: The Most Common Pitfalls!

Good quality starting material is king for reverse transcription! Obtaining reliable results in any experiment requires good preparation. We often take reverse transcription for granted, and we don’t always consider that our qPCR might be performing poorly because of problems in that step. Since it’s quite often the reverse transcription reaction itself that causes fuss […]

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In PCR, qPCR and qRT-PCR 19th of January, 2017
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The Obligate qPCR Standard Curve

At first sight, real-time PCR looks like a very simple technique—very straightforward. Also, when it’s optimized, real-time PCR leads to interesting results. However, to obtain consistent and accurate results reflecting the reality, good controls are crucial for SYBR qPCR.  One of these controls is the qPCR standard curve to check for the efficiency of your primers. Efficient Primers […]

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In PCR, qPCR and qRT-PCR 17th of November, 2016
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From Revolution to Evolution: Stem-loop Real-time PCR

Kary Mullis invented polymerase chain reaction (PCR) in 1985 creating a revolution in molecular biology techniques. But it hasn’t stopped there. PCR has greatly evolved over the years. Today, we stand at a point, where we can clone micro RNAs (miRNAs) in real time! Due to miRNA size (about 18-21 nucleotides long) and varied expression levels, […]

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In PCR, qPCR and qRT-PCR 27th of September, 2016
real-time PCR read on

The Real-Time PCR Digest

In the 30 odd years since its invention, the polymerase chain reaction (PCR) has become the bread and butter technique of molecular biologists. The secret to its indispensability lies in its simplicity and versatility. Numerous variants of the technique have been developed; one of these, real-time PCR, has become the method of choice for quantitative […]

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In PCR, qPCR and qRT-PCR 15th of September, 2016
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Methods for Relative Quantification of qPCR Data. Yes, There is More Than One.

As all of you probably know, methods for calculating relative gene expression from qPCR data include: a) double delta Ct (ΔΔCt) and b) that one other method. Chances are you’ve probably gotten beyond the ΔΔCt method, but you should be prepared in case you face primer sets of different amplification efficiencies. Both methods require the […]

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In PCR, qPCR and qRT-PCR 18th of August, 2016
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Decoding Cancer: Practical Advice for Working with Cellular Heterogeneity

In this webinar, you will learn about the process for genotyping cells—particularly tumor cells—and practical advice for analyzing the resulting data. The main points that we will cover are: Practical advice and methods for sample collection and preparation for your genotyping experiments An overview of the available data analysis methods for your genotyping results Examples […]

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In PCR, qPCR and qRT-PCR, Qiagen 9th of August, 2016 Presented by: Ania Wronski
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SPUD’s Your Bud When it Comes to Sensitive qPCR

There’s piloting a brand new technique for the first time. Then, there’s jumping through hoops trying to get an established lab technique to work. The former, in contrast to the latter, is expected to be fraught with hardships. Yet troubleshooting an old lab technique that isn’t working anymore, is frustrating at a whole new level. […]

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In PCR, qPCR and qRT-PCR 21st of July, 2016
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PCR problems read on

PCR Problems? Try an Additive

You’ve tried all the usual stuff, and checked the primer sequences twice, but still can’t get that PCR fragment amplified. It’s time to enter the strange world of PCR additives. Over the years a variety of additives have been shown to enhance PCR reactions in certain situations. Here is a summary of some of the […]

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In PCR, qPCR and qRT-PCR 9th of July, 2016