BMBMethod:PCI
<owwmenu font="arial, helvetica, sans-serif" bold="1" color="black" bgcolor="white" hovercolor="black"
bghovercolor="gray" topFontSize="10" fontSize="8" pagewidth="650" image="OldMain2.jpg" lab="BMBMethod">
Home=
Meetings=#,Schedule=Schedule, Past=Past, Presentations=Presentations
Protocols=Protocols, Protein=Protocols#Protein, DNA=Protocols#DNA, RNA=Protocols#RNA, Other=Protocols#Other
Tips & Tricks=TipsandTricks, Protein=TipsandTricks#Protein, DNA=TipsandTricks#DNA, RNA=TipsandTricks#RNA, Other=TipsandTricks#Other
Equipment=Equipment, Departmental=Equipment#BMBDepartment, Personal=Equipment#Personal, Other=Equipment#Other
Reagents=Reagents, Departmental=Reagents#Departmental, Cheap Buys=Reagents#Cheap Reagents
</owwmenu>
Phenol-Chloroform Extraction of DNA (PCI)
This technique can be used to remove or inactivate proteins (such as restriction enzymes or RNAse) from DNA preps.
- Start with sample volume between 50 and 400 ul in an eppendorf tube
- Add an equal volume of PCI (25:24:1 Phenol:Chloroform:Isoamyl Alcohol, pH 7.9) to your sample [Take from LOWER phase]
- Vortex thoroughly
- Centrifuge 2 min
- Remove upper phase into new tube, do not disturb interface.
- Upper phase will form bubble as its volume gets small
- Add equal vol of CI (24:1, Chloroform:Isoamyl Alcohol)
- Vortex
- Centrifuge 2 min
- Remove upper phase to new tube
- Perform ethanol precipitation to finish cleaning DNA