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W4502 Sigma

Water

Molecular Biology Reagent

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Preparation Note

This product is prepared from double distilled and deionized water

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Milli-Q Integral System
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Safety Information

Personal Protective Equipment 
WGK Germany 
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RTECS 
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Protocols & Articles

Articles

Qualitative Multiplex PCR Assay for Assessing DNA Quality from FFPE Tissues and Other Sources of Damaged DNA

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

Protocols

Animal Tissue DNA-Extraction & WGA Amplification Protocol

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...

Antibody-Enzyme Mediated Hot Start PCR Protocol

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

Blood Card - Extraction & Amplification WGA Protocol

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...

Buccal DNA Extraction & WGA Amplification Protocol

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...

Cloning the Gene-of-Interest into a Plasmid Vector

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

Complete Whole Transcriptome Amplification Kit WTA2 Protocol

Product Description Components Storage/Stability Procedure Troubleshooting Guide Frequently Asked Questions Materials References Publication

Experienced User Protocol | GenElut Mammalian Genomic DNA Miniprep Kit

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

Extraction & Amplification of whole blood using WGA-Protocol

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...

GenElute Bacterial Genomic DNA Kit Protocol

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: Amplification, Anticoagulants, Cell disruption, Centrifugation, Degradations, Digestions, Evaporation, Molecular biology, Nucleic acid denaturation, Phase transitions, Polymerase chain reaction, Precipitation

GenElute Bacterial Genomic DNA Kit, Experienced User Protocol

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

GenElute Mammalian Genomic DNA Miniprep Kit Protocol

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

GenElute Plant Genomic DNA Miniprep Kit Protocol (G2N10, G2N70, G2N350)

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

GenElute™ Blood Genomic DNA Kit Protocol (NA2010, NA2020)

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

GenomePlex Complete Whole Genome Amplification Kit

Product Description Components Storage Procedure References Troubleshooting Guide Frequently Asked Questions Disclaimer Automation Protocol Posters and Publications Reagents Required

GenomePlex Whole Genome Amplification Kit Protocol

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

GenomePlex® WGA Reamplification Kit Protocol (WGA3)

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...

Imprint Methylated DNA Quantification Protocol

Note: Storage Temperature - see Storage/Stability section for individual component storage conditions

Multiplex qPCR Protocol

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

MystiCq® MicroRNA® Quantitation System

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

Plant DNA extraction & WGA Amplification-protocol

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...

Plasma Serum DNA extraction & WGA Amplification Protocol

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 ...

Primer Concentration Optimization Protocol

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

Primer Optimization Using Temperature Gradient Protocol

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

Qualitative Multiplex PCR Assay for Assessing DNA Quality from FFPE Tissues and Other Sources of Damaged DNA

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, Applications, Cell disruption, Degradations, Forensic, Methods, Polymerase chain reaction, Polymerase chain reaction - quantitative, Sequences

Restriction Enzyme Cloning Manual Buffer Recipes

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

Reverse Transcription Protocol (One-step Probe Detection)

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

Reverse Transcription Protocol (One-step SYBR Green I Dye Detection)

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

Reverse Transcription Using ReadyScript cDNA Synthesis RDRT Protocol

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

SPUD Assay for Detection of Assay Inhibitors Protocol

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

SYBR® Green I Dye Quantitative PCR Protocol

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

Saliva DNA Extraction & WGA Amplification Protocol

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...

Selecting Correctly Expressing Recombinants

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

SeqPlex RNA Amplification Kit Protocol

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...

Standard Reverse Transcription Protocol (Two-step)

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' Assay for Analysis of RNA Integrity Protocol

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® Whole Transcriptome Amplification Protocol

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

WGA4 - GenomePlex Single Cell Whole Genome Amplification Kit

Product Description Reagents Provided Storage/Stability Procedure References Troubleshooting Guide FAQs Posters and Publications Downstream applications         Gel Electrophoresis & qPCR         CGH...

qPCR Efficiency Determination Protocol

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

qPCR Gene Expression/Copy Number Analysis Using SYBR Green I Dye Detection Protocol

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

qPCR Gene Expression/Copy Number/SNP Analysis Using Probe Detection Protocol

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

qPCR Reference Gene Selection Protocol

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

qPCR Using a Single Detection Probe Protocol

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

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