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…be designed to amplify the target DNA sequence. Although PCR-ELISA is unlikely to replace conventional ELISA or Real-Time PCR (see Real-Time PCR article) in the detection of proteins, biomolecules and…
…quantification in real-time RT-PCR. Nucleic Acids Res. 29:e45. Lin CH et al. (2011) Quantification bias caused by plasmid DNA conformation in quantitative real-time PCR assay. PLoS One. 6:e29101 Karlen, Y…
…product. Whenever you flick open a tube of amplified PCR product, or pipette PCR product, a small amounts of your PCR product becomes aerosolized. This aerosolized PCR product or amplicon…
…quantitative real-time PCR experiments) guidelines recommend using Cq value. These guidelines standardize experimentation and reporting, which is critical given that quantitative real-time PCR is used to analyze clinical samples and…
…MassTag PCR? MassTag PCR is a type of multiplex PCR that uses mass spectrometry to distinguish between PCR products. To recap briefly, multiplex PCR allows you to detect multiple targets simultaneously in…
…preferential amplification of low levels of mutated DNA, whereas conventional PCR amplifies all sequences indiscriminately. Colony PCR: Small quantities of bacterial cells from bacterial colonies are added to the PCR…
…the DNA polymerase amplifying the wrong template DNA sequence, is one cause of non-specific bands on gel following PCR. How to Eliminate Non-Specific Amplification in PCR Careful design of PCR…
…behaves as an individual PCR reaction and, as with real-time PCR, fluorescent probes are used to identify amplified target DNA. At the end of PCR, each partition is readily analyzed…
…controls, can make troubleshooting easier. But Is UNG Any Good for Real-Time PCR? However, as more and more labs are moving into real-time PCR analysis for quantification purposes, I wonder…
…to a 3D printer to build it. Designed to provide real-time PCR capabilities out in the field or other resource-limiting settings. [1] Arduino PCR Thermocycler $85 The parts needed to…
…and avoid it in the future? What is PCR contamination? The biggest source of PCR contamination is aerosolized PCR products, which are created when you open a tube or pipette amplified…
There are many different methods and protocols on making your PCR run more efficiently. I recently came across an interesting PCR method called “nanoparticle” PCR. This method seems to attract…
…miRNAs and other small RNAs”. Advantages to Stem-loop Real-time PCR Overall, the stem-loop real-time PCR technique provides the following advantages: The technique is highly specific for miRNAs, and nonspecific genomic…
…pathogen loads in clinical samples, and detecting SNPs/mutations. What is Real-time PCR? Real-time PCR, as the name suggests, measures the amount of PCR product at the end of each amplification…
…Estimating PCR efficiency from the slope of an amplification curve so you don’t need to run standard curves in qPCR Looking for new PCR reagents and equipment PCR troubleshooting and…
… Nested PCR Touchdown PCR and Stepdown PCR Multiplex PCR General PCR troubleshooting guide Useful References Chapter 3 Quantitative real-time PCR (qPCR) Hydrolysis (Taqman) probes…
So you’re designing a new experiment that requires PCR quantification. You used to have only one method to choose from, but now you have two – Quantitative Real-Time PCR (qPCR)…
…the magic happens. All of the above ingredients are added to a PCR tube and the tube is thermocycled. To achieve thermocycling when PCR was first invented, individual PCR tubes…
Nowadays, almost every biology lab has a PCR instrument — from portable, battery-operated machines, to “PCR-by-water baths”, do-it-yourself PCR, or familiar vendor packages, including those with real-time quantification or droplet…
…you learn about photonic PCR. What the Heck is Photonic PCR “So what is it?” Well, the conventional PCR uses “hotplates” to regulate the temperature inside a reaction tube. It…
…am sold on two-step, rather than one-step, qRT-PCR. Are you? References: 1. Spelman, F et al, Analytical Biochemistry, 303, 95-98 (2002). 2. Sigma Aldrich Technical Guide to quantitative PCR: https://www.sigmaaldrich.com/life-science/molecular-biology/pcr/quantitative-pcr/qpcr-technical-guide.html…
…for colony PCR fall into two categories: Insert-specific primers Backbone-specific primers Because many cloning projects begin by PCR amplifying an “insert”, many recommend using these same primers for colony PCR….
…ALL. THE. TIME.) called “real time PCR” and shortened to “RT-PCR”. Oh, biologists! RT-qPCR, or reverse-transcriptase quantitative PCR, is, as you probably guessed, a combination of the two methods mentioned…
…of Quantitative real-time PCR Experiments (MIQE). A recently published update now provides a checklist for digital PCR – Minimum Information for Publication of Quantitative Digital PCR Experiments (dMIQE). MIQE checklist…
…PCR may be your answer! In this article, we will discuss what it is, its benefits, and how to do overhang PCR in the lab. What Is Overhang PCR? Overhang…
…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 inside of each partition. Upon…
…dried PCR ingredients that don’t require refrigeration. Greener PCR Solar PCR is designed for medical diagnostic personnel in resource-limited areas. By negating the need for a reliable flow of clean electricity,…
…Alu-PCR steps in. Alu-gag nested PCR coupled with real time PCR for quantitation Unlike the previous Alu-PCR protocols, the newest method, Alu–gag nested PCR, incorporates 2 steps of PCR to…
…following insertion of the PCR product. Using topoisomerase kits means that you no longer need to design restriction sites into your PCR primers. This is great if your PCR product…
…the PCR product. A Simple Solution: Nested PCR Nested PCR is a truly elegant solution. It uses two pairs of primers: the first set bind your target sequence but rather…
…could have a major effect on PCR efficiency. 6. Customer Support What is the current expertise of the people involved in real-time PCR? If you have a lot of new…
Are you even a real researcher if you have never cried over a qRT-PCR result? The biggest challenge in setting up a quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) is ensuring…
…the PCR project was back in the oven and over the next few years it was perfected and various applications developed, including DNA fingerprinting (1986), gene amplification systems (1988), real-time…
…the amplified DNA is run on a gel and stained with ethidium bromide. Real-time PCR (rtPCR), also known as quantitative PCR (qPCR), instead correlates production of the PCR product to…
…solve this problem and help you get a seamless PCR. Need help making your PCR fail (a bit) less often? Download our free notorious PCR inhibitors poster and pin it…
What do bunnies, coins and PCR have in common? They all have tails! (ha ha!) Thermal asymmetric interlaced PCR or TAIL-PCR is used to sequence and analyse unknown DNA fragments…
…worthwhile. One such technique is PCR cloning. This article is intended to help you decide if PCR cloning is for you. What is PCR Cloning? PCR cloning is a method…
…choice for you by comparing Digitial PCR with Quantitative Real-Time PCR. Recommended References Huggett, JM and Whale, A (2013) Digital PCR as a Novel Technology and Its Potential Implications for Molecular…
…a comment below. References Gaudet M, Fara AG, Beritognolo I, Sabatti M. (2009) Allele specific PCR in SNP Genotyping. Methods Mol Biol. 578:415–24. Abbon, M. Get that Genotyping PCR to Work…
…the best results. Using Touchdown PCR One of my favorite tricks is the touchdown PCR method, in which your first few PCR rounds are at a higher annealing temperature. After…
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