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Molecular Beacons
Molecular Beacons are structured probes that are highly sensitive, sequence specific, and are used for sequence detection in qPCR and in vitro studies.
Dual-Labeled Probes
Dual-Labeled Probes are the most common probe type for qPCR and are often referred to as hydrolysis probes.
Troubleshooting PCR and RT-PCR Amplification
This page shows PCR and RT-PCR amplification troubleshooting.
How qPCR Works
Watch these videos to learn how real time or quantitative PCR (qPCR) works and the benefits of both the SYBR Green-based and probe-based methods of qPCR assay.
Universal ProbeLibrary System Technology
UPL is based on only 165 short hydrolysis probes. They are labeled at the 5' end with fluorescein (FAM) and at the 3' end with a dark quencher dye.
Quantitative PCR and Digital PCR Detection Methods
Fluorescent dyes or probes are included in PCR mixes to monitor the change in NA amplicon concentration as the reaction proceeds.
RNA Immunoprecipitation (RIP) FAQs
RNA Immunoprecipitation (RIP) Frequently Asked Questions
Probe-based qPCR
Probe based QPCR utilizes a fluorescent–labeled target-specific probe resulting in increased specificity and sensitivity. Additionally, a variety of fluorescent dyes are available so that multiple primers can be used to simultaneously amplify many sequences.
Multiplex qPCR with JumpStart™ Taq ReadyMix™ for Quantitative PCR
Method outlines use of a hot start Taq for multiplex qPCR and provides guidance on how to optimize dNTPs, primer, probes and MgCL2 concentrations. By optimizing these parameters, the user can improve assay sensitivity and linear range of detection.
PCR/qPCR Data Analysis
After a traditional PCR has been completed, the PCR/qPCR data analysis is conducted by resolution through an agarose gel or, more recently, through a capillary.
Using Probe-Based Quantitative PCR (Qpcr) to Measure Gene-Level Expression
Quantitative PCR (qPCR) provides information about gene expression, gene amplification or loss, and small alterations. qPCR is often used to investigate tumor biology and to discover the genetic and epigenetic causes of cancer
LNA Probes
Locked Nucleic Acid is a novel type of nucleic acid analog that contains a 2'-O, 4'-C methylene bridge.
Technical Guide to PCR Technologies
Examples of basic PCR/qPCR/dPCR protocols that can be used as the foundation for explorations into some of the concepts described in the theoretical chapters of this guide. Included are detailed protocols for assay quality control, in addition to more general...
Oligonucleotide Quality Control by Mass Spectrometry
Mass Spectrometry is the technology of choice for analyzing oligonucleotide synthesis. It enables the most sensitive detection of low levels of by-products, which can affect performance.
PCR-based Assay Regulations and Validation
While many PCR assays are developed for research applications there are further considerations for those that are being developed to become diagnostic assays or to be performed in support of: Biologics License Application (BLA), New Drug Application (NDA), Premarket Approval...
PCR Assay Optimization and Validation
PCR assay guide navigates you through primer validation and other assay optimization factors to ensure high sensitivity and specificity for optimum DNA/ RNA quantification.
Direct and Crude Extract qPCR
Rapid qPCR analysis from highly inhibited tissue and blood sample extractions with KAPA PROBE
Array CGH Analysis of Challenging Samples
In recent years, array-based Comparative Genomic Hybridization (aCGH) has been refined to determine chromosomal changes at progressively higher resolutions. This evolving technology is, however, hampered by the large DNA input requirement—a minimum of 150,000 copies of a human genome, or...
Oligonucleotide Quantification
DNA absorbs ultraviolet light due to its highly conjµgated nature. DNA may thus be easily quantitated in a UV spectrometer.
Dual-Labeled DNA Probes: Fluorescent Probes
To achieve accurate and reliable results from your real-time quantitative PCR, choose from the broadest range of sequence–specific, fluorescent probes: LightCycler probes, Dual-labeled fluorogenic probes, Molecular Beacons, Scorpions™ probes. All of our fluorescent probes are deprotected, desalted and purified...
Primer and Probe Design
Video: Primer and Probe Design
DNA Oligonucleotide Synthesis
Learn about the steps in phosphoramidite solid-phase method of Oligonucleotide Synthesis method that we use for DNA oligonucleotide manufacturing.
PCR/qPCR/dPCR Assay Design
The entire PCR workflow is vulnerable to factors which introduce variability. Many of the variable components are unavoidable, such as the source of the sample or the requirement for a reverse transcription step. Assay design is also highly variable and...
Complete Solutions for PCR Assay Development
Fit-for-use products offer the quality, consistency & documentation necessary for every step of your IVD development and manufacturing process.
Mgat4 May Play a Role in Increased Sialylation by Overexpressing Functional MGAT1 in Mgat1-Disrupted Chinese Hamster Ovary (CHO) Cells
MGAT1 adds N-acetylglucosamine to the Man5GlcNAc2 (Man5) structure. Goh et al. reported increased sialylation after restoring MGAT1 function in MGAT1 deficient CHO cells.
KiCqStart® SYBR® Green Primers FAQ
KiCqStart™ Primers Frequently Asked Questions; Sigma-Aldrich.com
Choosing the Right Custom qPCR Probe
There is a wide selection of possible custom probes that can be used for qPCR, and each has advantages for different applications.
RT-PCR / RT-qPCR Troubleshooting
Developing a PCR or RT-PCR/RT-qPCR troubleshooting protocol so that data are reliable is essential. Potential sources of RT-PCR or PCR error and problems include operator error, the PCR master mix, and oligo design. This PCR troubleshooting guide outlines and details...
OligoArchitect™ Assay Design
Efficient qPCR relies on good assay design. Since the invention of PCR, many parameters have been identified as important for assay quality, such as estimates of oligo temperature characteristics, GC content and folding properties.
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