CHLOROFORM: ISOAMYL ALCOHOL (24:1)

Specifications

Chloroform: Isoamyl Alcohol 24:1 denatures proteins, facilitates the separation of organic and aqueous phases, and reduces foaming during the extraction process. It is highly volatile, toxic, and flammable at storage temperatures around 85°C; direct contact should be minimized. Commonly used in DNA/RNA extraction via phase separation method.

Chloroform: Isoamyl Alcohol 24:1

Manual instruction

Chloroform: Isoamyl alcohol 24:1 is used to separate the lysate mixture into two distinct phases, with the upper aqueous phase containing DNA/RNA. Below is an example of a DNA/RNA extraction procedure utilizing Chloroform: Isoamyl alcohol 24:1:

• Grind 5 g of plant tissue using a mortar and pestle in liquid nitrogen.
• Add 500 μL of CTAB buffer and mix thoroughly.
• Incubate the sample at 65°C for 30 minutes.
• Add 200 μL of Chloroform: Isoamyl alcohol 24:1 and mix vigorously.
• Centrifuge at 13,000 rpm for 5 minutes. Carefully transfer 500 μL of the upper aqueous phase to a new tube.
• Add an equal volume of ice-cold absolute isopropanol (2–8°C) and incubate at –20°C for 10 minutes.
• Centrifuge at 13,000 rpm for 10 minutes.
• Discard the supernatant, wash the pellet with 500 μL of 70% ethanol, and allow the DNA to air dry completely.
• Resuspend the DNA in 50 μL of TE buffer (10 mM Tris, 1 mM EDTA, pH 8.0)

Read more:

• Other Bio-reagents
• Manual instruction