Phenol:Chloroform:Isoamyl Alcohol 25:24:1 Saturated with 10 mM Tris, pH 8.0, 1 mM EDTA, 1 X 400 mL (P2069-400ML)

For applications requiring a higher pH, such as the isolation of large intact genomic DNA, addition of the Equilibration Buffer is recommended. After the addition of Equilibration Buffer, the product is stable for 6 months vs. 2 years without added buffer. Storage at -20 °C protected from light extends the shelf life to greater than 1 year after the addition of Equilibration Buffer. Add the entire contents of the small bottle of Equilibration Buffer to the large bottle of Phenol:Chloroform:Isoamyl Alcohol. Mix gently and allow the phases to separate before use, approximately 2-4 hours. This will adjust the pH of the Phenol solution from pH 6.7 ± 0.2 to 8.0 ± 0.2. This lot of phenol:chloroform:isoamyl alcohol 25:24:1 was use tested by extracting a crude preparation of pBR322 DNA preparation from E. coli. Based on evaluation by agarose gel electrophoresis the extracted DNA was not degraded or reduced in concentration. The same procedure was applied to purified pBR322 DNA and lambda Hind III digest DNA. Based on electrophoretic evaluation the purified preparations were not detectably degraded or reduced in concentration.

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