Search results for: real-time PCR
Polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA)
…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…
Read MoreThe Devil is in the Details: How to Setup a PCR Laboratory
…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…
Read More11 qPCR Papers Every Researcher Should Know
…assay design, troubleshooting, and data analysis. qPCR (real-time PCR) protocol explained Recent and brief YouTube video. A perfect introduction if you are completely new to qPCR. MIQE qPCR & dPCR…
Read MoreAn Introduction to MassTag PCR
…know that PCR’s main purpose is to amplify a stretch of nucleic acids based on sequence-specific primers. Nowadays, a wide range of PCR techniques exist, including qPCR, RACE-PCR, Digital PCR,…
Read MoreThe A-Z of PCR Variants
…the vector flanking the insertion site, resulting in amplification of the inserted sequence. Digital PCR (dPCR): This style of PCR allows for more accurate quantification of nucleic acid amounts. Rather…
Read MoreWhat Is a Cq (Ct) Value?
…of quantitative real-time PCR experiments) guidelines recommend using Cq value. Therefore throughout this article, we will use Cq only. [1] What is the Cq Value? Real-time PCR (often called qPCR)…
Read MoreEntering the Digital Age for Quantitative PCR Analysis: Digital PCR
…PCR success! Quantitative PCR (qPCR) is probably one of the most important PCR spin-offs to date. As the name suggests, qPCR allows you to quantify DNA, and it is not…
Read MoreClean Up Your Act! How To Clean Up PCR Contamination
…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…
Read MoreSmall Particles (Things) Matter!- Introducing Nanoparticle PCR
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…
Read MoreSkip DNA Extraction and Go Direct to PCR with VersaTaq TM Direct PCR Polymerase
…is resistant to many potent PCR inhibitors. This mutant Taq jump starts your setup directly to the PCR step, eliminating costly extraction steps. With the VersaTaq Direct PCR Polymerase protocol,…
Read MoreTop 10 websites to help you with your PCR experiments
…everything you ever wanted to know about qPCR, including emerging qPCR applications, chemistries, methods and algorithms behind qPCR, Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE), cyclers, kits,…
Read MoreUnderstand and troubleshoot PCR with The BitesizeBio Guide to PCR
…Bands appearing in negative controls Alternative PCR methods Nested PCR Touchdown PCR and Stepdown PCR Multiplex PCR General PCR troubleshooting guide Useful References Chapter 3 Quantitative real-time PCR (qPCR) Hydrolysis…
Read MoreTaking the Pain, Challenge and Rage out of PCR: The BitesizeBio Guide to PCR
…PCR runs and delves into more complex considerations that are essential for qPCR and digital PCR (dPCR) users. See the below excerpt for a example of what this book includes…
Read MoreThe Real-Time PCR Digest
…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…
Read MoreDigital PCR or Quantitative Real-Time PCR: Which Method Is Best for Your Quantitative PCR Application?
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)…
Read MoreAmplify Your PCR Success with the Right PCR Instrument!
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…
Read MorePhotonic PCR: When Lightening Strikes Your DNA
…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…
Read MoreWhat is PCR? The Beginner’s Guide
PCR is THE technique of modern molecular biology labs. If you need to copy, sequence, or quantify DNA, you need to know PCR. But how do you get started with…
Read MoreDecisions, Decisions: How to Choose the Best qPCR Probe for Your Experiment
…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…
Read MoreMIQE Guidelines for Digital PCR
…for publication of real-time PCR experiments (qPCR) suggested by SA Bustin et al. in 2009. MIQE guidelines for qPCR and dPCR publications are divided into the following categories: essential information…
Read MoreDivide and Conquer: How to Setup Your First Droplet Digital PCR Experiment
…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…
Read MoreHow to Be Greener – The Environmentally Friendly Guide to PCR
Science is an expensive business and those who use high energy-demanding techniques may not even realize just how expensive they are. The Cost of PCR Let’s looks at PCR. You…
Read MoreHow to Quantify Integrated HIV Genomes Using Alu-gag PCR
…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…
Read MorePCR on a shoestring: Build Your Own PCR Machine
…A few awesome folks have come up with ways of making a PCR machine from scratch and made the information open-source, including OpenPCR ($649), the Coffee Cup PCR ($350), Arduino PCR…
Read MoreProtocols for Cloning Without Restriction Enzymes or Ligases
…vector 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…
Read MoreTime to Instigate Nested PCR
…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…
Read MoreThe History of PCR
…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…
Read MoreThermal Asymmetric Interlaced PCR (TAIL-PCR)
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…
Read MoreShould PCR cloning be a part of your molecular cloning toolkit?
…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…
Read MoreOverhang PCR
…realize you wanted to tag it with something? Or you just want to add restriction enzymes to your PCR product to make it easier to clone into a plasmid? Overhang…
Read MoreTouchdown PCR: A Primer and Some Tips
…this article I’ll provide a primer on touchdown PCR (TD-PCR) and some tips and references for perfecting it. So let’s start with… What is touchdown PCR? TD-PCR is a modification…
Read MoreWhat is Digital PCR?
…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…
Read MoreQuantifying Allele-Specific Gene Expression Using PCR-Based Methods
…you can selectively amplify both alleles in a single PCR. Allele-Specific Gene Expression Using qPCR With access to qPCR reagents and equipment, you can take a more quantitative approach to…
Read MoreFaster PCR Optimization
…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…
Read MoreNo More White Elephants! – Consider this Before Buying a Real-time PCR Cycler
…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…
Read MoreThe Inhibitors Haunting Your PCR
…to the PCR Gods. And still, something unknown messes up your results. This unknown at work is generally a PCR inhibitor. Before you blame it all on Murphy’s law and…
Read MorePCR Pitfalls: The Devil is in the Details
PCR was actually one of the first lab techniques I learned as an undergrad. Despite being sometimes labeled as a pretty basic lab skill, PCR doesn’t always work as expected….
Read MoreGet that Genotyping PCR to Work EVERY TIME
…performed through genotyping PCR. What is Genotyping PCR? Genotyping PCR is used to determine the genotype of an organism (e.g., WT vs. mutant, or WT vs. transgenic). PCR primers are…
Read MoreMediator Probe PCR
…mediator sequence results in dequenching of the UR fluorophore by release of the quencher or strand displacement. Mediator Probe PCR works with both DNA and RNA targets in real-time PCR…
Read MoreDetecting Signal in qPCR: From DNA Binding Dyes to BHQ Probes
…really began with the use of a camera to detect the generation of fluorescence in ‘real-time’ as the PCR cycling was progressing. The theory worked on the simple basis that…
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