PHENOL: CHLOROFORM: ISOAMYL ALCOHOL (25:24:1)

Specifications

Phenol: Chloroform: Isoamyl Alcohol 25:24:1 is used in the purification of nucleic acids. The product consists of chloroform, isoamyl alcohol, and phenol saturated at pH 7.9.

It is a homogeneous, transparent, and volatile solution, ready for immediate use.

The solution is packaged in amber-colored bottles and should be stored at 2–8°C.°C.

Phenol:Chloroform:Isoamyl Alcohol 25:24:1 – 100ml Bottle

Manual instruction

Reference protocol:

• Prepare 800 μl of lysed cell sample in a 2 ml microcentrifuge tube. Add 400 μl of Phenol saturated pH 7.9, mix thoroughly, and centrifuge at 12,000 rpm for 1 minute.
• Collect the upper aqueous phase, add 800 μl of Phenol: Chloroform: Isoamyl alcohol 25:24:1 , mix well, and centrifuge at 12,000 rpm for 1 minute.
• Collect the upper aqueous phase, add 700 μl of Chloroform: Isoamyl alcohol 24:1, mix thoroughly, and centrifuge at 12,000 rpm for 1 minute.
• Collect the upper aqueous phase, then add 1/10 volume of 3 M Sodium Acetate and 2 volumes of cold 96–100% ethanol
• Mix gently until a precipitate forms. Centrifuge at 12,000 rpm, discard the supernatant, and wash the DNA pellet with 70% ethanol. Perform a minimum of 2 and up to 5 washes. After the final wash, remove residual ethanol and dry the pellet in a 50°C incubator for 15 minutes.
• Add 500 μl of 1X TE buffer. Incubate the tube in a water bath at 65–70°C for 30 minutes, or until the DNA is fully dissolved.

Caution

• Phenol is highly toxic upon contact with skin, eyes, ingestion, or inhalation. It may cause chemical burns.
• In case of skin contact, wash immediately with soap and plenty of clean water. Remove any contaminated clothing promptly.
• If you feel unwell, seek medical attention immediately.
• Do not use expired products.

Read more:

• Other Bio-reagents
• Manual instruction