Search results for: real-time PCR
Mediator 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 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 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…
Read MoreProblems Amplifying GC-rich regions? 5 Easy Solutions
…PCR requires the addition of 7-deaza-2′-deoxyguanosine, a dGTP analog to the PCR mixture. Slow-down PCR also uses a standardized cycling protocol with varying temperatures. Generally, ramp rates are lowered, and…
Read MoreOld Reliable: Two-Step Allelic Exchange
…upstream (PCR #1) and downstream (PCR #2) of my gene of interest and then PCR those two fragments together for a clean deletion in the SOE process (PCR #3). The…
Read MoreHow to Survive a Difficult PCR
…witches. Well, scientific witches—by which I mean: valid, solid reasons, for why a PCR is not working! It is so frustrating to spend time and energy on a PCR, and…
Read MoreJust What Do All These Additives Do?
…While the above PCR additives can improve PCR results, it is impossible to predict which additives at what concentrations are best for your PCR needs. Therefore, like always, you need…
Read MoreNeed to clean up PCR products? Now there’s a faster solution with HT ExoSAP-IT® Fast Reagent!
…is a better way to clean your PCR products! USB® products from Affymetrix®, the leader in enzymatic PCR clean up, has released a faster and more stable version of its…
Read MoreLetting Go of Your Ligase: Transfer PCR
…yet. PCR is like this sweater – it goes with almost everything and molecular biology is taking full advantage of this using it at every chance it gets. Since PCR…
Read MoreOverlap Extension PCR Cloning
…it works The first of two PCR reactions allows you to create a linear insert containing plasmid sequence at both ends. These will enable the strands of the PCR product…
Read MorePCR vs qPCR Sensitivity
…more sensitive? References 1Experts Roundtable: Real-time PCR and microbiology. In Real-time PCR in microbiology: from diagnosis to characterisation? ed: Ian M. Mackay. Caister Academic Press 2J. Clin. Microbiol. 2008 Jun;46(6):1897-900….
Read MoreTo boil? Or be boiled? Saving Time With Colony PCR
…can directly perform PCR using the colonies on the petri dish that the post-doc left on your bench. Some people refer to it as “Colony PCR” while others call it,…
Read MorePolymerase Incomplete Primer Extension (PIPE) Cloning Method
…incomplete extension products and it’s these guys that make PIPE PCR possible. By following certain primers design rules, the PCR process introduces short, overlapping sequences at the ends of these…
Read MoreSome Sanger Sequencing Tips and Tricks
…specific PCR primers, simply add the M13 sequences to the 5′ ends. Order these from your oligo provider and your PCR is (almost) ready to sequence. PCR clean-up: You need…
Read MoreDesigning Luck: 8 Basic Concepts for Designing Primers for a Standard PCR
I think we all have been through those my-PCR-product-didn’t-get-amplified days. Sometimes, playing around a bit more with the PCR conditions brings luck, or sometimes it doesn’t work at all. These…
Read More10 Handy Tips to Help Keep Your PCRs Contamination Free
…example, keep tubes and strips for PCR in a separate container and use those only for PCR. If possible have a different freezer/fridge location for pre-PCR reagents and post-PCR reactions….
Read MoreThe Essential PCR Troubleshooting Checklist
Routine PCR? Let’s be honest, there’s no such thing. Even with the simplest PCR reaction things can go wrong, so you need to have a good checklist of ideas for…
Read MoreGet To Know Your DNA Polymerases
While most may think standard Taq is the backbone of PCR, many other DNA polymerase options exist. The polymerase you use significantly impacts the efficacy of your PCR, specifically on…
Read MoreA Quick Tour Around Probe-based Multiplexing qPCR
…By relying on fluorescent dyes or fluorophore-tagged probes to measure the amount of amplified product, it eliminates the need for gel electrophoresis. Instead, real-time PCR instruments can measure the fluorescence…
Read MoreImportant Considerations for Determining qPCR Efficiency
…and pain later. For a more comprehensive guide to PCR, download our free PCR fundamentals eBook and become an expert. More reading on related topics Ten Tips for Consistent Real-time PCR…
Read MoreFinding Nemo: Understanding Single Cell Isolation and PCR Amplification
…AmpliGrid, you will need an AmpliSpeed slide cycler. Two of the most commonly used options are: RT-PCR: reverse transcriptase enzyme is used during the PCR to convert RNA to complementary…
Read MoreHow to Screen for CpG Methylation by Methylation Specific PCR
…done by performing methylation-specific PCR on bisulfite-converted DNA. Primer design For methylation-specific PCR you include CpGs in the primer sequences. That means you have to choose whether you want to…
Read MoreThe Invention of PCR
…breakthrough, together with advances in oligonucleotide synthesis made PCR both cost effective and convenient and it quickly entered mainstream research. As researchers flocked to PCR, improvements to and variations on…
Read MoreChoosing the Correct Enzyme for PCR
A wide variety of enzymes are available for PCR and RT-PCR and the optimal choice depends on a range of factors specific to your experiment. Some of these factors will…
Read MoreqPCR: RNA Quality and Why It Matters
Gene expression analysis plays a pivotal role in a wide range of studies, including biomedical analysis and diagnostics. Of all the methods available for gene expression analysis, quantitative real-time PCR…
Read MoreMultiple Fragment Ligation: The Why and How
…Here is a brief list to help you along. Ligation Reagent Functions PCR or synthetic oligonucleotides Standard PCR reagents and oligo design techniques are used to amplify your desired fragments. …
Read MoreAnchored Multiplex PCR for Next Generation Sequencing
…methods can be difficult, which is why Anchored Multiplex PCR is so great. Another strength of Multiplex PCR is its ability to enrich small and poor quantities of nucleic acid….
Read MoreA Beginner’s Guide to How Blunt-End Cloning Works
…circular construct, their absence can also be used to your advantage. If you are using PCR to create the insert or to amplify a linearized vector, PCR primers lacking 5’-phosphates…
Read MoreSingle Molecule Real-Time Sequencing
…technology is that you obtain real-time sequencing results. Today, I’m going to talk about a different NGS platform that also allows real-time sequencing, but it uses a different principle. Sounds…
Read MoreApps That Bring PCR to Your Mobile Device
…free version which can be upgraded for extra capability. Life technologies also have a general Real-Time PCR app which contains a reaction calculator, troubleshooting guide amongst other feature. This is…
Read MoreHow to scale up a PCR reaction
…amount of PCR product? The Mix I’ll start from the premise that you have already optimised your PCR reaction, you know the best conditions and have established the PCR cannot…
Read MoreControl your error! How to minimize pipetting error and get low SDs in qRT-PCR
…performance of your reagents. For quantitative real time polymerase chain reaction (qRT-PCR) there may be several factors that influence technical replicates including slight differences caused by plate variability, or freeze…
Read MoreHow to Train a Junior Colleague in the Basics of PCR
…on PCR. You don’t want to lead him/her astray, but it’s hard to remember the parts that you struggled with in your early days. This article will remind you of…
Read MoreLet’s Get Melting
What is HRM Analysis? It is now more than a decade since the introduction of melting analysis to characterize PCR products. Melting analysis following SYBR green-based real-time PCR has become…
Read MoreA Molecular Biology Roadmap For First Time Travelers
…Well in molecular biology, all roads lead to PCR– and away from PCR– and back to PCR again. Basically, PCR is the central technique in all of molecular biology, and…
Read MoreSix Steps to Successful Mouse Genotyping
…For all further characterizations of offspring from established mouse lines, use PCR, because it’s quick and easy. You can use a conventional end-point PCR with an agarose gel readout for…
Read MoreThe Art of PCR Primer Design
…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 for…
Read MorePCR 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….
Read MoreDo-it-Yourself PCR
…that the basic technology to perform PCR is affordably and openly available to all. In the past one of the main barriers for introducing PCR technology at small labs, as…
Read MoreHow Are Crimes Solved by PCR?
…hair particles, they can attempt to identify the killer using PCR and genetic fingerprinting, providing the first solid lead towards solving this crime. So Where Does PCR Come into All…
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