P2069

Sigma-Aldrich

Phenol:Chloroform:Isoamyl Alcohol 25:24:1 Saturated with 10 mM Tris, pH 8.0, 1 mM EDTA

Supplied with Equilibration buffer, for molecular biology

NACRES:
NA.52
Pricing and availability is not currently available.

Quality Level

grade

for molecular biology

pH range

6.5 - 6.9 (pH of phenolic phase)
7.8 - 8.2 (pH of phenolic phase after the addition of Equilibration Buffer)

Featured Industry

Agriculture

storage temp.

2-8°C

Application

Phenol:Chloroform:Isoamyl Alcohol has been used in the preparation of RNA for the study of total RNA isolation from myocardial biopsies. It has been also used to extract DNA during Human Androgen Receptor (HUMARA) Clonality Assay.
For applications requiring a higher pH, such as the isolation of large intact genomic DNA, addition of the Equilibration Buffer is recommended.

Packaging

100, 400 mL in glass bottle
Supplied with P3803 and a separate bottle of equilibration buffer to adjust the phenol phase to pH 8.0 ± 0.2.

Suitability

This lot of phenol:chloroform:isoamyl alcohol 25:24:1 was use tested by extracting a crude preparation of pBR322 DNA preparation from E. coli. Based on evaluation by agarose gel electrophoresis the extracted DNA was not degraded or reduced in concentration. The same procedure was applied to purified pBR322 DNA and lambda Hind III digest DNA. Based on electrophoretic evaluation the purified preparations were not detectably degraded or reduced in concentration.

Preparation Note

Add the entire contents of the small bottle of Equilibration Buffer to the large bottle of Phenol:Chloroform:Isoamyl Alcohol. Mix gently and allow the phases to separate before use, approximately 2-4 hours. This will adjust the pH of the Phenol solution from pH 6.7 ± 0.2 to 8.0 ± 0.2.

Other Notes

After the addition of Equilibration Buffer, the product is stable for 6 months vs. 2 years without added buffer. Storage at -20 °C protected from light extends the shelf life to greater than 1 year after the addition of Equilibration Buffer.

Signal Word

Danger

Target Organs

Central nervous system

RIDADR

UN 2810 6.1 / PGIII

WGK Germany

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

  1. Which document(s) contains shelf-life or expiration date information for a given product?

    If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.

  2. How do I get lot-specific information or a Certificate of Analysis?

    The lot specific COA document can be found by entering the lot number above under the "Documents" section.

  3. There are two layers in the bottle of Product P2069, Phenol:Chloroform:Isoamyl Alcohol 25:24:1; which layer should I use?

    The lower phase is the phase that is to be used.

  4. There is an Equilibration Buffer (Product B5658) that is included with Product P2069, Phenol:Chloroform:Isoamyl Alcohol 25:24:1.  Is B5658 also available separately?

    No, it is only available as part of P2069.

  5. I ordered Product P2069, Phenol:Chloroform:Isoamyl Alcohol 25:24:1; I got one bottle of the Phenol:Chloroform:Isoamyl Alcohol and one bottle of B5658, Equilibration buffer. When and how am I supposed to use the B5658?

    B5658 is a Tris buffer that is used to raise the pH of the phenol:chloroform:isoamyl alcohol mixture, to make it suitable for isolation of genomic DNA. The contents of the two bottles need to be well mixed. Then allow the mixture to separate; this may take 2-4 hours. This procedure should raise the pH of the phenol phase from 6.7 to about 8.0.  Only one "treatment" is needed. This solution should be stable for up to 12 months.

  6. How do I find price and availability?

    There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote.  USA customers:  1-800-325-3010 or view local office numbers.

  7. What is the Department of Transportation shipping information for this product?

    Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product. 

  8. I see that you list two equilibrated Phenol:Chloroform:Isoamyl alcohol products. When should I use Product P3803, and when should I use Product P2069?

    In short, P3803 is for RNA work, and P2069 is for DNA work. Product P2069 consists of one bottle of Product P3803 and one bottle of Product B5658. The B5658 is added to the P3803 to raise the pH of the organic phase from 6.7 to 8.0; pH 8.0 is not suitable for RNA work, because RNA can degrade at the higher pH.

  9. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Aaron M New et al.
Nature communications, 10(1), 3657-3657 (2019-08-16)
We lack an understanding of how the full range of genetic variants that occur in individuals can interact. To address this shortcoming, here we combine diverse mutations between genes in a model regulatory network, the galactose (GAL) switch of budding...
Yongzhong Wang et al.
Microcirculation (New York, N.Y. : 1994), 13(3), 187-197 (2006-04-22)
Angiogenic therapies in animals have demonstrated the development of new blood vessels within ischemic myocardium. However, results from clinical protein and gene angiogenic trials have been less impressive. The present study aimed to investigate the expression of angiogenic genes in...
Martin Häring et al.
Frontiers in behavioral neuroscience, 9, 235-235 (2015-09-22)
The endocannabinoid (eCB) system possesses neuromodulatory functions by influencing the release of various neurotransmitters, including γ-aminobutyric acid (GABA) and glutamate. A functional interaction between eCBs and the serotonergic system has already been suggested. Previously, we showed that cannabinoid type-1 (CB1)...
Amber L Scott et al.
Molecular biology and evolution, 34(10), 2690-2703 (2017-09-29)
Polyploidization events have occurred during the evolution of many fungi, plant, and animal species and are thought to contribute to speciation and tumorigenesis, however little is known about how ploidy level contributes to adaptation at the molecular level. Here we...
Laura Cook et al.
Cellular and molecular gastroenterology and hepatology, 9(3), 485-506 (2019-12-04)
Bacterial flagellin is an important antigen in inflammatory bowel disease, but the role of flagellin-specific CD4+ T cells in disease pathogenesis remains unclear. Also unknown is how changes in intestinal microbiome intersect with those in microbiota-specific CD4+ T cells. We...
Protocols
Method for reverse transcription of RNA into DNA. Uses a premixed reagent that contains reverse transcriptase, dNTPs, primers, RNase inhibitor and buffer. Fast generation of cDNA.
Read More

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service

Social Media

LinkedIn icon
Twitter icon
Facebook Icon
Instagram Icon

MilliporeSigma

Research. Development. Production.

We are a leading supplier to the global Life Science industry with solutions and services for research, biotechnology development and production, and pharmaceutical drug therapy development and production.

© 2021 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.

Reproduction of any materials from the site is strictly forbidden without permission.