3 Purification of Nucleic Acids From Eukaryotic Cells
65
Materials
• cells or tissue
• chloroform
• isoamyl alcohol
• phenol (water-saturated)
• ß-mercaptoethanol
• 0.5M EDTA (pH8.0)
• ethanol
• guanidinium thiocyanate
• 2M sodium acetate (pH 4.0)
• 1 M sodium citrate (pH 7.0)
• 10% (w/v) sodium lauryl sarkosinate
• 70% and 100% ethanol, ice-cold
• RNAse free TE-buffer (pH 7.0) (10 mM Tris-HCl (pH 7.0), 1 mM EDTA
(pH 8.0))
• DEPC-treated water
Reagents
• 4 to 15 ml disposable polypropylene tubes (e.g. Falcon, Cat.Nos. 2063, Supplies
2059)
• sterilized microcentrifuge tubes
• microcentrifuge at 4°C
• chemical hood
• spectrometer
Procedure
1. In advance prepare the following solution:
- 49:1 (v/v) chloroform:isoamyl alcohol
- solution Da:
ln advance
65
Materials
• cells or tissue
• chloroform
• isoamyl alcohol
• phenol (water-saturated)
• ß-mercaptoethanol
• 0.5M EDTA (pH8.0)
• ethanol
• guanidinium thiocyanate
• 2M sodium acetate (pH 4.0)
• 1 M sodium citrate (pH 7.0)
• 10% (w/v) sodium lauryl sarkosinate
• 70% and 100% ethanol, ice-cold
• RNAse free TE-buffer (pH 7.0) (10 mM Tris-HCl (pH 7.0), 1 mM EDTA
(pH 8.0))
• DEPC-treated water
Reagents
• 4 to 15 ml disposable polypropylene tubes (e.g. Falcon, Cat.Nos. 2063, Supplies
2059)
• sterilized microcentrifuge tubes
• microcentrifuge at 4°C
• chemical hood
• spectrometer
Procedure
1. In advance prepare the following solution:
- 49:1 (v/v) chloroform:isoamyl alcohol
- solution Da:
ln advance