Invitrogen™ Acid-Phenol:Chloroform, pH 4.5 (with IAA, 125:24:1)

Product Code.	Quantity	unitSize
10544645	400 mL	Each
10095904	100 mL	Each

Product Code. 10544645 Supplier Invitrogen™ Supplier No. AM9722

Description

In RNA extraction procedures, Acid Phenol:Chloroform:IAA aids in the removal of DNA (it partitions into the organic phase), helps to stabilize the interface, and prevents foaming when mixing.

Acid Phenol:Chloroform:IAA (125:24:1) is premixed and supplied at pH 4.5 ± 0.2. Provided in one bottle of 400 mL. In RNA extraction procedures, Acid Phenol:Chloroform:IAA aids in the removal of DNA (it partitions into the organic phase), helps to stabilize the interface, and prevents foaming when mixing. Preparation of phenol for use in molecular biology applications is a time-consuming and often hazardous procedure due to its toxic and corrosive nature. Ambion premixed, quality-tested, saturated phenols are ready to use, eliminating handling problems and offering a more convenient, safer, and easier alternative to preparing solutions from crystalline phenol.

Order Info

Shipping Condition: Room temperature

Specifications

• Chemical Name or Material: Phenols
• Content And Storage: Store at 4°C or -20°C.
• Packaging Type: Bottle
• pH: 4.5
• Product Line: Ambion
• Purity: Molecular Biology Grade
• Quantity: 400 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.

Safety and Handling

Product Identifier

• ACID PHENOL:CHLOROFORM 400 ML

Signal Word

• Danger

Hazard Category

• Acute toxicity Category 3
• LONG-TERM AQUATIC HAZARD Chronic 2
• Carcinogenicity Category 2
• Serious eye damage/eye irritation Category 1
• Germ cell mutagenicity Category 2
• Reproductive toxicity Category 2
• Skin corrosion/irritation Category 1 A
• Specific target organ toxicity Category 1

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.
• H351-Suspected of causing cancer.
• H361-Suspected of damaging fertility or the unborn child.
• H372-Causes damage to organs.
• H411-Toxic to aquatic life with long lasting effects.

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, chloroform

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