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Phenol Equilibration

  Phenol Equilibration
1. -20°C에 보관하고 있는 액상 phenol을 꺼내어 상온으로 항온한다.
2. 68°C 수조에서 phenol을 녹인다.
3. 산화방지제인 hydroxyquinoline을 최종 농도 0.1%가 되게 첨가하고 잘 섞는다.
4. 동량의 0.5 M Tris-Cl buffer (pH 8.0)를 첨가하고 15 분 동안 강하게 교반한다.
5. 교반을 중지하고 두 개의 층으로 분리되면 상층(완충액 층)의 용액을 aspiration으로 최대한 제거한다.
6. 동량의 0.1 M Tris-Cl buffer (pH 8.0)를 첨가하고 15 분 동안 강하게 교반한다.
7. 교반을 중지하고 두 개의 층으로 분리되면 상층(완충액 층)의 용액을 aspiration으로 최대한 제거한 후
   pH paper로 하층(phenol 층)의 pH를 측정한다.
8. 하층의 pH가 7.8 이상이 될 때까지 "6." ~ "7." 과정을 반복한다.
9. 하층의 pH가 7.8 이상이 되면 1/10 부피의 0.1 M Tris-Cl buffer (pH 8.0, 0.2% β-mercaptoethanol 포함)를 첨가한다.
10. 차광하여 4°C에서 보관한다.
Notes : Phenol은 부식성(corrosive)이 매우 강하므로 phenol을 다룰 때는 반드시 비닐장갑, 마스크, 보호용 안경을
착용하고 chemical hood에서 작업한다. 피부에 닿았을 경우에는 충분한 물로 씻어주되 ethanol로 씻지 않는다.


Technical Tables for Molecular Biology
Acrylamide Gel Electrophoresis of Xylene Cyanole FF and Bromophen..
Acrylamide Gel Electrophoresis of Proteins
Agarose Gel Electrophoresis of Xylene Cyanole FF and Bromophenol ..
Amino Acids
Amino Acids
Antibiotic Solutions
Genetic Markers
Genotypes
Nucleic Acid Data
pH of 50 mM Tris-HCl Buffer Solutions
Physical Constants of the Nucleoside Triphosphates
Recommended Gel Percentages for Separation of Linear DNA
Specifications of Common Vectors


Enzymes for DNA and RNA Modification
DNA Polymerases
Ligases
Nucleases
Phosphatases
Reverse Transcriptases
RNA Polymerases
Others


Protocols
DNA Mini-Agarose Gel Electrophoresis
Transformation of E. coli (CaCl2 and heat-shock method)
Mini-Prep of Plasmid DNA (Alkaline Lysis Method)


Preparation of Liquid solutions and Buffers
45% Acrylamide Solution
Actinomycin D Solution
0.1 M Adenosine 5'-Triphosphate (ATP) Solution
10 M Ammonium Acetate Solution
10% Ammonium Persulfate (APS) Solution
20 mg/ml 5-Bromo-4-Chloro-3-Indolyl-β-D-Galactopyranoside (X-Gal..
50 mg/ml 5-Bromo-4-Chloro-3-Indolyl Phosphate (BCIP) Solution
1 M CaCl2 Solution
50 mM Deoxyribonucleoside Triphosphate (dNTP) Solutions (Each)
1 M Dithiothreitol (DTT) Solution
10 mg/ml Ethidium Bromide (EtBr) Solution
0.5 M Ethylenediaminetetraacetic Acid (EDTA) Solution, pH 8.0
200 mg/ml Isopropylthio-β-D-Galactoside (IPTG) Solution
50 mg/ml Nitro Blue Tetrazolium (NBT) Solution
Phenol Equilibration
Phosphate-Buffered Saline (PBS) Solution
Potassium Acetate Solution(For Alkaline Lysis Method of Plasmid P..
20X Saline-Sodium Citrate (SSC) Buffer
20X Saline-Sodium Phosphate-EDTA (SSPE) Buffer
3 M Sodium Acetate (pH 5.2 and pH 7.0) Solution
20% Sodium Dodecyl Sulfate (SDS, Lauryl Sulfate Sodium Salt) Solu..
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