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
…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…
Read MoreAn Introduction to MassTag PCR
…is 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…
Read MoreThe A-Z of PCR Variants
…PCR. This is used for single cell genetic diagnosis and the identification of high threat bio agents, as it is more efficient and highly specific than conventional PCR. Long-PCR: Using…
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 MoreWhat is Touchdown PCR? A Primer and 5 Easy Tips For Success
…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…
Read MoreEntering the Digital Age for Quantitative PCR Analysis: Digital 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…
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 MoreHow to Build Your Own PCR Machine
…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…
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 MoreEliminate PCR Amplicon (but not real-time PCR?) Carry-Over With UNG
…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…
Read MoreTop 10 websites to help you with your PCR experiments
…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…
Read MoreUnderstand and Troubleshoot PCR with The BitesizeBio Guide to PCR
… 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…
Read MoreWhat is PCR? The Beginner’s Guide
…making your PCR fail (a bit) less often? Download our free notorious PCR inhibitors poster and pin it up near your DNA engine. Or download the Bitesize Bio PCR eBook…
Read MoreTaking the Pain, Challenge and Rage out of PCR: The BitesizeBio Guide to PCR
…PCR methods such as nested PCR or touchdown PCR may be more successful. How to use this e-book You can use this PCR guide to trouble-shoot where your gel electrophoresis…
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 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
…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…
Read MoreHow to Be Greener – The Environmentally Friendly Guide to PCR
…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…
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 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 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 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 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 MoreThe Inhibitors Haunting Your PCR
…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…
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
…PCR may be your answer! What is it? Overhang PCR is a technique that utilizes the intrinsic fidelity of the 3’ end of primers for a specific sequence to enable…
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
…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…
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 MoreDo You Know Where You Are? Find Out with Genome Walking
…is just a fancy way of saying “doing a PCR reaction on the amplicon of an earlier PCR reaction” or “a PCR within a PCR”. This second nested PCR will…
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…
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