DNA isolation from mammalian samples.

DNA isolation from mammalian samples.

Current protocols in molecular biology (2013-04-03)

Ximeng Liu, Shuko Harada

ABSTRACT

DNA isolation is a fundamental initial step for molecular genetic studies. Two quite different methodologies are described in this unit: silica spin column and phenol/chloroform extraction. Currently, the most commonly used technique is the silica-based spin column, although phenol/chloroform extraction is still widely used. This unit also presents basic procedures for extraction of DNA from both fresh tissues and formalin-fixed, paraffin-embedded (FFPE) tissue samples.

MATERIALS

Product Number

Brand

Product Description

472476

Sigma-Aldrich

Chloroform, contains amylenes as stabilizer, ACS reagent, ≥99.8%

77608

Sigma-Aldrich

Phenol, BioUltra, for molecular biology, ≥99.5% (GC)

16018

Sigma-Aldrich

Phenol, ≥96.0% (calc. on dry substance, T)

242322

Sigma-Aldrich

Phenol, contains hypophosphorous as stabilizer, loose crystals, ACS reagent, ≥99.0%

W322318

Sigma-Aldrich

Phenol, ≥99%

33517

Sigma-Aldrich

Phenol, puriss. p.a., ACS reagent, reag. Ph. Eur., 99.0-100.5%

P1037

Sigma-Aldrich

Phenol, for molecular biology

P5566

Sigma-Aldrich

Phenol, BioXtra, ≥99.5% (GC)

185450

Sigma-Aldrich

Phenol, unstabilized, ReagentPlus^®, ≥99%

40063

Supelco

Phenol solution, 5000 μg/mL in methanol, certified reference material

328111

Sigma-Aldrich

Phenol, unstabilized, purified by redistillation, ≥99%

02487

Supelco

Chloroform, analytical standard

P4682

Sigma-Aldrich

Phenol solution, Saturated with 0.1 M citrate buffer, pH 4.3 ± 0.2, BioReagent, for molecular biology

35952

Supelco

Phenol, PESTANAL^®, analytical standard

P4557

Sigma-Aldrich

Phenol solution, Equilibrated with 10 mM Tris HCl, pH 8.0, 1 mM EDTA, BioReagent, for molecular biology

288306

Sigma-Aldrich

Chloroform, anhydrous, ≥99%, contains 0.5-1.0% ethanol as stabilizer

16016

Sigma-Aldrich

Phenol, puriss., ≥99.5% (GC), meets analytical specification of Ph. Eur., BP, USP, crystalline (detached)

C7559

Sigma-Aldrich

Chloroform, ≥99%, PCR Reagent, contains amylenes as stabilizer

77607

Sigma-Aldrich

Phenol, BioUltra, for molecular biology, TE-saturated, ~73% (T)

46344

Supelco

Phenol solution, 100 μg/mL in acetonitrile, PESTANAL^®, analytical standard

C2432

Sigma-Aldrich

Chloroform, contains 100-200 ppm amylenes as stabilizer, ≥99.5%

PHR1047

Supelco

Phenol, Pharmaceutical Secondary Standard; Certified Reference Material

16017

Sigma-Aldrich

Phenol, puriss., meets analytical specification of Ph. Eur., BP, USP, 99.5-100.5% (GC)

P9346

Sigma-Aldrich

Liquified Phenol, ≥89.0%

372978

Sigma-Aldrich

Chloroform, anhydrous, contains amylenes as stabilizer, ≥99%

366919

Sigma-Aldrich

Chloroform, ACS spectrophotometric grade, ≥99.8%, contains 0.5-1.0% ethanol as stabilizer

319988

Sigma-Aldrich

Chloroform, contains ethanol as stabilizer, ACS reagent, ≥99.8%

48688

Supelco

Phenol solution, certified reference material, 500 μg/mL in methanol