Invitrogen™ UltraPure™ Buffer-Saturated Phenol

Product Code.	Quantity	unitSize
11598886	100 mL	100mL
11508896	400mL	400mL

Product Code. 11598886 Supplier Invitrogen™ Supplier No. 15513039

Description

UltraPure™ Buffer-Saturated Phenol is used in the purification of nucleic acids. The reagent, which consists of UltraPure™ Phenol that has been saturated with Tris-HCl buffer, is already buffer equilibrated to pH >7.4.

• UltraPure™ Buffer-Saturated Phenol is used in the purification of nucleic acids
• The reagent, which consists of UltraPure™ Phenol that has been saturated with Tris-HCl buffer, is already buffer equilibrated to pH >7.4
• When mixtures are extracted with UltraPure™ Buffer- Saturated Phenol, proteins are denatured and collect in the organic phase or at the interphase, while most nucleic acids remain in the aqueous phase
• UltraPure™ Buffer-Saturated Phenol contains no preservatives
• It is packaged under an inert gas in shatter-resistant, plastic-coated amber bottles
• The appearance of the solution is evaluated at room temperature
• No RNase or DNase activity detected
• Store in refrigerator at 2°C to 8°C

Order Info

Shipping Condition: Room temperature

Specifications

• Chemical Name or Material: Phenols
• Content And Storage: Store in refrigerator (2–8°C).
• Packaging Type: Bottle
• pH: 7.4
• Product Line: UltraPure
• Purity: Molecular Biology Grade
• Quantity: 100 mL
• Shipping Condition: Room Temperature
• Form: Liquid

Frequently Asked Questions (FAQs)

Do you have any information on DNA and RNA purification using phenol chloroform and alcohol precipitation?

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.

What is the recommended protocol for phenol-extraction removal of proteins from nucleic acid containing solutions?

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.

What is the optimal pH of the phenol:chloroform mixture for isolation of DNA?

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.

Recently I came across a DNA purification technique, which uses urea during phenol extraction. What is the purpose of using urea?

Using urea during phenol extraction denatures the protein associated with the DNA and the proteins that bind the genomic DNA to the cell wall.

Safety and Handling

Product Identifier

• Buffer-Saturated Phenol

Signal Word

• Danger

Hazard Category

• Acute toxicity Category 3
• Serious eye damage/eye irritation Category 1
• Germ cell mutagenicity Category 2
• Skin corrosion/irritation Category 1 B
• Specific target organ toxicity after repeated exposure Category 2

Hazard Statement

• H301-Toxic if swallowed.
• H311-Toxic in contact with skin.
• H314-Causes severe skin burns and eye damage.
• H331-Toxic if inhaled.
• H341-Suspected of causing genetic defects.
• H373-May cause damage to organs.

Precautionary Statement

• P201-Obtain special instructions before use.
• P260-Do not breathe dust/fume/gas/mist/vapours/spray.
• P261-Avoid breathing dust/fume/gas/mist/vapours/spray.
• P264-Wash hands thoroughly after handling.
• P280-Wear protective gloves/protective clothing/eye protection/face protection.
• P301+P310-IF SWALLOWED: Immediately call a POISON CENTER/doctor /
• P302+P352-IF ON SKIN: Wash with plenty of water/soap.
• P303+P361+P353-IF ON SKIN (or hair): Take off immediately all contaminated clothing. Rinse skin with water/ shower.
• P304+P340-IF INHALED: Remove person to fresh air and keep comfortable for breathing.
• P305+P351+P338-IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.
• P403+P233-Store in a well-ventilated place. Keep container tightly closed.
• P501b-Dispose of contents/container in accordance with local/regional/national/international regulations.

Supplemental information

• MIXTURE LIST-Contain: phenol

For Research Use Only. Not for use in diagnostic procedures.