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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...
Polymerase Chain Reaction
The polymerase chain reaction is one of the most widely used techniques in molecular biology. The PCR process consists of three main steps, Denaturation, Annealing & Extension
Precision Genetic Modifications
Our scientists’ years of experience and Sigma-Aldrich’s access to multiple advanced gene editing technologies allow us to offer a cell engineering service with unparalleled success. We can engineer a variety of modifications in your cell line of choice to address...
Whole Transcriptome Amplification of RNA from Low Cell-Number Samples
The efficacy of amplification of small quantities of total RNA with the Complete Whole Transcriptome Amplification Kit (WTA2) was examined in this study.
5'/3' RACE Kit, 2nd Generation Troubleshooting
In addition to the troubleshooting provided in the product manual, most probably the efficiency of the tailing reaction performed by Terminal Transferase could be impaired. This could occur due to several reasons (which will not only affect the control reaction...
ThermaStop™-RT PCR Additive
PCR additive interacts with reverse transcriptase at low temperatures to reduce priming problems that lead to nonspecific bands.
KAPA2G Fast PCR Kits FAQ
Fast PCR is PCR in which total reaction time is decreased by reducing the duration of one or more of the steps during each cycle of a PCR. Using the KAPA2G Fast PCR Kit, PCR time can be reduced by...
A highly efficient method to concentrate DNA for forensic STR genotyping using DNAstable®
Forensic laboratories routinely use STR genotyping for identity testing of biological samples. However, forensic samples often contain low copy numbers of target DNA, making it difficult to obtain complete STR profiles.
Modification of the WTA2 Amplification Product for Next Generation Sequencing
Transplex Whole Transcriptome Amplification (WTA2) exponentially amplifies RNA producing a double-stranded cDNA library while precisely maintaining differential levels of individual transcripts in test and reference samples.
Restorase® DNA Polymerase
Restorase® was developed for researchers unable to achieve amplification of damaged DNA templates when using other commercially available DNA polymerases.
KAPA2G Fast Multiplex Kits FAQs
Frequently asked questions (FAQs) for KAPA2G Fast Multiplex Kits
Troubleshooting PCR and RT-PCR Amplification
This page shows PCR and RT-PCR amplification troubleshooting.
What is Hot Start PCR?
Learn about the benefits of using Hot Start PCR reagents to suppress enzymatic activity before PCR reaction tubes enter the thermal cycler.
Hot Start PCR
The purpose of Hot Start PCR is to inhibit the PCR reaction in order to reduce nonspecific amplification, prevent the formation of primer dimers, and increase product yields.
Dual-Labeled Probes in Digital PCR
Digital PCR (dPCR) is an end-point PCR method that is used for absolute quantification and for analysis of minority sequences against a background of similar majority sequences.
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.
FastStart™ Taq DNA Polymerase, 5 U/μL Protocol & Troubleshooting
The choice of the PCR enzyme in combination with an appropriate buffer can profoundly affect PCR outcome.
Extract-N-Amp™ Plant Tissue PCR Kits
The Extract-N-Amp™ kits are designed to rapidly extract and amplify genomic DNA. The plant tissue version of these kits has been optimized to amplify without concern over plant inhibitors. This technical document will discuss the versions of this kit that...
DNA Oligonucleotide Synthesis
Learn about the steps in phosphoramidite solid-phase method of Oligonucleotide Synthesis method that we use for DNA oligonucleotide manufacturing.
Retinoic Acid and Gene Expression
All-trans retinoic acid (RA, ATRA) is a pleiotropic activation factor that regulates genes associated with normal vertebrate cellular processes such as cell differentiation, cell proliferation, apoptosis, and embryonic development.
Oligonucleotide Quality Control & Quality Assurance
The foundation of Sigma's manufacturing process is a robust Quality Management System, which drives compliance to ISO 9001:2008 certification.
Mycoplasma PCR ELISA Protocol
Mycoplasmas are potential contaminants in mammalian cell culture manufacturing. All products produced in cell culture to be tested for the presence of Mycoplasma
PCR Master Mix
A PCR master mix is a batch of PCR or RT-PCR reagents that can be divided among many PCR reaction tubes. It usually includes DNA polymerase, dNTPs, MgCl2 and buffer. Make your own master mix or choose a commercial one.
KAPA Express Extract Kit FAQs
Frequently asked questions (FAQs) for KAPA Express Extract Kit.
KAPA2G Robust PCR Kits FAQs
Frequently asked questions (FAQs) for KAPA2G Robust PCR Kits.
Oligonucleotide Array CGH Analysis of a Robust Whole Genome Amplification Method
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, somewhat hampered by the large DNA input requirement—a minimum of 150,000 copies of a human genome...
Extract-N-Amp™ Blood PCR Kit Protocol
The Extract-N-Amp Blood PCR Kits contain all the reagents needed to rapidly extract and amplify human genomic DNA from whole blood, whole blood dried on a blood card, and cultured mammalian cells.
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...
Roche PCR Reagents and PCR Protocols
Roche polymerase chain reaction (PCR) reagents for reducing the error rate of Taq DNA polymerase and PCR selection guide.