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  • P2069 - Phenol:Chloroform:Isoamyl Alcohol 25:24:1 Saturated with 10 mM Tris, pH 8.0, 1 mM EDTA

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P2069 Sigma

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

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Properties

Related Categories Acids and Bases, Biochemicals and Reagents, Core Bioreagents, DNA & RNA Purification, Denaturation,
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

Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

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.

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.

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.

Application

For applications requiring a higher pH, such as the isolation of large intact genomic DNA, addition of the Equilibration Buffer is recommended.

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.

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.

Price and Availability


LC-MS Grade Solvents and Reagents

Microcon Centrifugal Filters

All labs need water
Safety & Documentation

Safety Information

Signal word 
Danger
RIDADR 
UN 2810 6.1 / PGIII
WGK Germany 
3

Frequently Asked Questions

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. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
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.
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.
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.
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. 
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, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
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.
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Protocols & Articles

Protocols

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

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