Water

While several genome editing tools have been developed in recent years, including zinc finger-based strategies and TALENs (transcription activator-like effector nucleases), none have been as efficien...
Keywords: Amplification, Cell culture, Enzyme-linked immunosorbent assay, Gene expression, Genetic, Genetics, Immunohistochemistry, Immunology, Nutrition, Polymerase chain reaction, Recombination, Southern blot, Transcription, Transfection, Western blot

*Calculations provide estimated results based on Antoine's Coefficients and their associated temperature range values. Antoine's Coefficients were obtained from the National Institute of Standards an...

The assessment of DNA quality is a crucial first step in acquiring meaningful data from formalin-fixed paraffin-embedded (FFPE) tissues, and other sources of damaged DNA. Using intact genomic DNA is ...
Steve Michalik, Christopher Williams
LSI Edition 23
Keywords: AGE, Amplification, Cancer, Cell disruption, Clinical, Degradations, Forensic, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Whole genome amplification

Animal tissue is a common source of material when performing genetic analysis. The protocol below is a simple method of extracting DNA from the animal sample. Once the DNA has been isolated, it can t...

During PCR assay preparation, nonspecific amplification can occur due to binding of PCR primers to nonspecific templates and from formation of primer dimers which result from using other primer molec...
Keywords: AGE, Amplification, Buffers, Electrophoresis, Enzyme activity, Gel electrophoresis, Nucleic acid denaturation, Polymerase chain reaction, Size-exclusion chromatography

The assessment of DNA quality is a crucial first step in acquiring meaningful data from formalin-fixed paraffin-embedded (FFPE) tissues, and other sources of damaged DNA. Using intact genomic DNA is ...
Keywords: AGE, Cell disruption, Degradations, Forensic, Polymerase chain reaction, Polymerase chain reaction - quantitative

Blood cards provide the convenience of archiving small volumes of blood. However, many times genomic DNA from these samples is limited, which may hinder the researcher’s ability to perform downstream...

This protocol provides a simple and convenient method to isolate, amplify, and purify genomic DNA from buccal swabs. Buccal swabs are a convenient method of acquiring a DNA sample. Once the DNA is is...

Genetic engineering is used in thousands of laboratories around the world. Given its importance it is remarkable that cloning strategies for many of the popular DNA components are not standardised. C...
Keywords: Antibiotics, Buffers, Cloning, Degradations, Dialysis, Digestions, Gene expression, Genetic, Genetics, Individual protein Immunoprecipitation, Molecular biology, Nucleic acid annealing, Peptide synthesis, Phosphorylations, Polymerase chain reaction, Precipitation, Purification, Sequencing, Substitutions, transformation

Product Description Components Storage/Stability Procedure Troubleshooting Guide Frequently Asked Questions Materials References Publication
Keywords: Amplification, Buffers, Centrifugation, Cloning, Condensations, DNA purification, Degradations, Microarray Analysis, Molecular probes, Nucleic acid annealing, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymerization reactions, Purification

For experienced users, an abbreviated protocol is listed below. For data, images, troubleshooting and other information, please see the full length protocol page.
Keywords: PAGE

Whole blood is a common source of material used to perform genetic analysis. Many times genomic DNA isolated from whole blood samples is of low yield. This can hinder the researcher’s ability to perf...

Sigma's GenElute™ Bacterial Genomic DNA Kit provides a simple and convenient way to isolate pure DNA from a variety of cultured bacteria. The kit contains all of the reagents needed to isolate and pu...
Keywords: AGE, Amplification, Cell disruption, Centrifugation, Degradations, Digestions, Electrophoresis, Evaporation, Gel electrophoresis, Molecular biology, Nucleic acid denaturation, Phase transitions, Polymerase chain reaction, Precipitation, Pulsed field gel electrophoresis

For experienced users, an abbreviated protocol is listed below. For data, images, troubleshooting and other information.

Sigma’s GenElute Mammalian Genomic DNA Miniprep Kit provides a simple and convenient way to isolate pure genomic DNA from a variety of cultured cells, tissues (including rodent tails), and fresh whol...
Keywords: AGE, Amplification, Anticoagulants, Cell disruption, Centrifugation, DNA purification, Degradations, Digestions, Electrophoresis, Evaporation, Gel electrophoresis, Molecular biology, Nucleic acid denaturation, Phase transitions, Polymerase chain reaction, Precipitation, Purification, Sample preparations, Sequencing

Several micrograms of DNA can be obtained from up to 100 mg of fresh tissue or 10 mg of freeze-dried material in less than an hour. The purified DNA is greater than 20 kb in length and can be used in...
Keywords: AGE, Amplification, Cell disruption, Centrifugation, Electrophoresis, Evaporation, Filtration, Gel electrophoresis, Molecular biology, Phase transitions, Polymerase chain reaction, Precipitation, Purification

Sigma’s GenElute™ Blood Genomic DNA Kit provides a simple and convenient way to isolate pure genomic DNA from fresh or aged (older than 24 hours) whole blood. The kit combines the advantages of silic...
Keywords: AGE, Addition reactions, Amplification, Anticoagulants, Cell disruption, Centrifugation, Digestions, Electrophoresis, Evaporation, Gel electrophoresis, Molecular biology, Nucleic acid denaturation, Phase transitions, Polymerase chain reaction, Precipitation, Purification, Sequencing

Product Description Components Storage Procedure References Troubleshooting Guide Frequently Asked Questions Disclaimer Automation Protocol Posters and Publications Reagents Required
Keywords: Absorption, Amplification, Centrifugation, Diagnostic, Dialysis, Genomics, Molecular biology, Molecular probes, Nucleic acid denaturation, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, Sequencing, Whole genome amplification

Product Description Components Storage/Stability Procedure Troubleshooting Guide Frequently Asked Questions Precautions and Disclaimer Materials References
Keywords: Absorption, Amplification, Centrifugation, Diagnostic, Dialysis, Genomics, Molecular biology, Molecular probes, Nucleic acid denaturation, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, Whole genome amplification

The GenomePlex WGA Reamplification Kit allows subsequent reamplifications of the WGA product with little genetic bias.  This kit contains an optimized enzyme that decreases the background in the reac...
Keywords: Amplification, Diagnostic, Dialysis, Genetic, Genomics, Molecular probes, Nucleic acid denaturation, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, Whole genome amplification

Note: Storage Temperature - see Storage/Stability section for individual component storage conditions
Keywords: Cardiovascular, Degradations, Diseases, Gene expression, Methylations, Molecular biology

In the PCR Technologies Guide, the requisite components and quality control requirements for qPCR experiments were described in detail. With those in mind, the following is a protocol that can be use...
Keywords: Nucleic acid annealing, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography

Technology Overview Workflow Background Protocols miRNA isolation cDNA synthesis miRNA qPCR Advanced information Product selection guides Troubleshooting
Keywords: AGE, Amplification, Buffers, Cell disruption, Centrifugation, Degradations, Diagnostic, Digestions, Diseases, Electrophoresis, Gel electrophoresis, Gene expression, Homogenization, Microarray Analysis, Molecular biology, Nucleic acid annealing, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Precipitation, Purification, RNA purification, Sample preparations, Separation, Solvents, Titrations

Extracting DNA from plant tissue is a complicated process due to the tough cell wall that surrounds most plant cells. Genomic DNA from plant material can be damaged during the extraction process, res...

Whole genome amplification (WGA) of plasma and serum DNA presents a unique challenge due to the small amount of nucleic acid in such samples. One must use a robust DNA purification scheme capable of ...

Optimization of qPCR conditions is important for the development of a robust assay. Indications of poor optimization are a lack of reproducibility between replicates as well as inefficient and insens...
Keywords: Nucleic acid annealing, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography

One approach to assay optimization is to determine the optimum annealing temperature (Ta) of the primers by testing identical reactions containing a fixed primer concentration, across a range of anne...
Keywords: Amplification, Nucleic acid annealing, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography

Induced pluripotent stem cells (iPSCs), have the capacity to give rise to differentiated progeny arising from of all germ layers of the body including: ectoderm, endoderm, and mesoderm. The ability t...
Keywords: Alzheimer Disease, Apoptosis, Cell culture, Centrifugation, Cloning, Culture media, Gene expression, Genetic, Microscopy, Phase transitions, Transfection

Tip 1: DNA is quite stable in TE buffer at 4ºC. If stored in elution buffer or water then freezing at -20 ºC is advised.
Keywords: Phase transitions, Sequencing, Sterilizations

In some cases it is preferable to measure the target transcript directly, without preparing cDNA from the entire RNA sample. Such situations may include measurements on highly degraded RNA or when th...
Keywords: Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography, Transcription

In the example given below, the primer concentrations can be adjusted according to the results of optimization procedures (see Primer Concentration Optimization, Primer Optimization Using Temperature...
Keywords: Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography

Preparation Place components on ice. Mix, and then briefly centrifuge to collect contents at the bottom of the tube

The SPUD assay is one option for identification of inhibitors that may be present in RNA or DNA samples. The assay is particularly useful when a large number of samples are to be analyzed or when tar...
Keywords: Amplification, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography

Although quantitative PCR uses the same basic concept as traditional PCR, the reactions differ in that the amplicons are generally smaller and are detected indirectly using an additional dye or label...
Keywords: Gene expression, Nucleic acid annealing, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography

Whole Genome Amplification can be performed on DNA extracted in many ways. Sigma-Aldrich offers many products for DNA extraction including the GenElute™ Blood Genomic DNA Kit (NA2010), GenElute Mamma...

Blue / White colony screening is a strategy to quickly and easily distinguish between recombinant and non-recombinant colonies. It requires a special vector and a special strain of E. coli. It is par...
Keywords: AGE, Amplification, Antibiotics, Cell disruption, Centrifugation, Cloning, Degradations, Detergents, Diagnostic, Digestions, Electrophoresis, Gel electrophoresis, Molecular biology, Peptide synthesis, Phase transitions, Polymerase chain reaction, Precipitation, Purification, Sequencing, Transfection, transformation

The SeqPlex RNA Amplification kit provides a method for amplification of total RNA or isolated mRNA prior to entry into the workflows of the commonly used deep sequencing platforms. Microgram quantit...
Keywords: Amplification, Centrifugation, Condensations, Degradations, Electrophoresis, Eliminations, Enzyme activity, Gene expression, Genomics, Microarray Analysis, Molecular probes, Nucleic acid annealing, Nucleic acid denaturation, Peptide synthesis, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymerization reactions, Purification, Sample preparations, Sequencing, Sonication

Reverse transcription (RT) is the process of converting RNA to cDNA using a reverse transcriptase enzyme and dNTPs.
Keywords: Amplification, Polymerase chain reaction, Polymerase chain reaction - quantitative, Transcription

The 3’/5’ integrity assay is a potential first step in the identification of RNA degradation. The assay is particularly useful when a large number of samples are to be analyzed or when the degradatio...
Keywords: Amplification, Degradations, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography, Transcription

TransPlex, a Whole Transcriptome Amplification (WTA) method, allows for representative amplification of nanogram quantities of total RNA in less than 4 hours without 3'-bias. Microgram quantities of ...
Keywords: Amplification, Centrifugation, Cloning, Condensations, DNA purification, Gene expression, Genomics, Microarray Analysis, Molecular biology, Molecular probes, Nucleic acid annealing, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymerization reactions, Purification, Sequencing

Product Description Reagents Provided Storage/Stability Procedure References Troubleshooting Guide FAQs Posters and Publications Downstream applications         Gel Electrophoresis & qPCR         CGH...
Keywords: Amplification, Cell disruption, Centrifugation, Degradations, Diagnostic, Electrophoresis, Gel electrophoresis, Genomics, Microarray Analysis, Molecular biology, Molecular probes, Nucleic acid denaturation, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymorphisms, Purification, Whole genome amplification

Once an assay has been optimized, it is important to verify the reaction efficiency. This information is important when reporting and comparing assays. In this example protocol, the assay efficiency ...
Keywords: Amplification, Gene expression, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography, Transcription

Measuring a target quantity relative to one or more stable reference genes using SYBR Green I dye detection is a common application of qPCR. Below is a standard protocol that can be adapted to specif...
Keywords: Nucleic acid annealing, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography, Transcription

The most common application for qPCR is the measurement of a gene transcript or copy number quantity relative to one or more reference genes using probe detection. The reactions may be designed such ...
Keywords: Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography, Transcription

Analysis of gene expression data requires a stable reference or loading control. This reference is usually one or more reference genes. Suitable reference genes are those which are unaffected by diff...
Keywords: Gene expression, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography, Transcription

In the PCR Technologies Guide, the requisite components and quality control requirements for qPCR experiments were described in detail. With those in mind, the following is a protocol that can be use...
Keywords: Nucleic acid annealing, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography

The Pressure-Temperature Nomograph tool is an application of the Clausius-Clapeyron Equation, which assumes the heat of vaporization is a constant over a pressure range. Antoine´s Equation gets aroun...
Keywords: Evaporation, Vaporization

a. Gant, R., LC/GC 10 (7), 515 (1992) 1. At 20 °C 2. At 20 °C +/- 5 °C relative to water at 4 °C 3. At 10 mmHg pressure 4. At l=589.32 nm
Keywords: Gas chromatography, High performance liquid chromatography