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Invitrogen™ UltraPure™ Phenol:Water (3.75:1, v/v)
Description
UltraPure Phenol:Water (3.75:1, v/v) is used in the purification of nucleic acids. This liquid reagent consists of UltraPure Phenol and highly pure deionized water. The reagent can be used to prepare different types of buffer-saturated phenol. When mixtures are extracted with buffer-saturated phenol, proteins are denatured and collect in the organic phase or at the interphase, while nucleic acids remain in the aqueous phase. UltraPure Phenol:Water (3.75:1, v/v) contains no preservatives. It is packaged under an inert gas in shatter-resistant, plastic-coated amber bottles.
Performance and Quality Testing
The appearance of the solution is evaluated at room temperature. No RNase or DNase activity detected.
Order Info
Shipping Condition: Room temperature
Specifications
Specifications
| Chemical Name or Material | Phenols |
| Content And Storage | Store in refrigerator (2–8°C). |
| For Use With (Application) | RNA/DNA Extraction |
| pH | 4.45 |
| Product Line | UltraPure |
| Purity | Molecular Biology Grade |
| Quantity | 400 mL |
| Shipping Condition | Room Temperature |
| Form | Liquid |
Frequently Asked Questions (FAQs)
Phenol extraction of proteins:
Phenol extraction is frequently used to remove proteins from nucleic acid solutions. A common protocol is to add an equal volume of buffer-saturated phenol or phenol:chloroform:isoamyl alcohol (25:24:1, v/v/v) to an aqueous nucleic acid solution, vortex, and centrifuge at 14,000 x g for 1 min to separate the phases.
Studies at Thermo Fisher Scientific have shown that the concentration of NaCl in the aqueous solution should not exceed 0.5 M for good recovery of DNA. Residual phenol can be removed from the aqueous phase by extraction with an equal volume of chloroform or ether. After extraction, DNA is usually precipitated with ammonium acetate and ethanol as described in another protocol on this server. Ref. Karger, B. D. (1989) FOCUS 11, 14.
A good source of general information on the properties of phenol can be found in Wallace, Donald M. “Large and Small-Scale Phenol Extractions”. Methods in Enz. Volume 152 guide to Molecular Cloning Techniques. 1987. Academic Press, Inc. Berger and Kimmel, eds. Chap.4, pg 33-41.
(a) At pH 5 to 6 DNA is selectively retained in the organic phase and interphase, leaving RNA in the aqueous phase. Therefore a pH greater than 7 is needed if DNA is to be extracted.
(b) At pH values below 7.6, poly A+ RNA is lost to the organic phase if chloroform is not present.
(c) Optimal RNA yields in phenol extraction are obtained if the salt concentration is less than 0.15 M NaCl. Salt concentration in the sample is not a factor for larger DNA molecules.
To store RNA after extraction use DEPC-treated water.
Below is a commonly used protocol:
(1) Add an equal volume of buffer-saturated phenol or phenol:chloroform:isoamyl alcohol (25:24:1, v/v/v) to an aqueous nucleic acid solution. Note: for RNA solutions, acid-phenol is recommended.
(2) Vortex, and centrifuge at 14,000 x g for 1 min to separate the phases.
(3) Residual phenol can be removed from the aqueous phase by extraction with an equal volume of chloroform or ether.
(4) After extraction, DNA/RNA is usually precipitated with ammonium acetate and ethanol.
Partitioning of the nucleic acids in phenol is pH dependent. At pH 7.0 or higher, both DNA and RNA partition into the aqueous phase. At an acidic pH (below 7.0) DNA is denatured and will move into the organic phase, but the RNA remains in the aqueous phase. The mixture should be adjusted to at least pH 7.4 for work with DNA.
Using urea during phenol extraction denatures the protein associated with the DNA and the proteins that bind the genomic DNA to the cell wall.
For research use only. Not for use in diagnostic procedures.
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