New Protocol for DNA Extraction of Stool

• Published in: BioTechniques Vol. 28; no. 2; pp. 286 - 290
• Main Authors: Machiels, B.M, Ruers, T, Lindhout, M, Hardy, K, Hlavaty, T, Bang, D.D, Somers, V.A.M.C, Baeten, C, von Meyenfeldt, M, Thunnissen, F.B.J.M
• Format: Journal Article
• Language: English
• Published: Natick, MA Future Science Ltd 01.02.2000; Eaton
• Subjects: Base Sequence; Biological and medical sciences; Biotechnology; Colorectal Neoplasms - diagnosis; Colorectal Neoplasms - genetics; Diverse techniques; DNA Primers - genetics; DNA, Neoplasm - genetics; DNA, Neoplasm - isolation & purification; Feces - chemistry; Feces - cytology; Fundamental and applied biological sciences. Psychology; Humans; Molecular and cellular biology; Molecular Probe Techniques; Polymerase Chain Reaction; Case study; Human; Polymerase chain reaction; DNA; Extraction; Feces; Method
• ISSN: 0736-6205; 1940-9818
• DOI: 10.2144/00282st05

Present methods for DNA isolation of stool have various limitations such as the amount of stool used, the requirement of lavage fluids or the use of fresh stool. In this paper, a new method is described for the isolation of human nucleic acids from stool, which is independent from the moment of collection. Fecal samples as dry as possible were collected from 75 patients; two grams of stool were mixed with a lysis buffer containing phenol. DNA yields of crude stool were variable and ranged from 9-1686 μg/g of feces. With dot blots in 9 of the 75 cases, the human DNA was identified and ranged from 0.06%-46%. In the remaining 66 cases, human genomic DNA was detected by nested PCR, using human K- gene amplification as an example. Amplification products were confirmed for human K- with the exonuclease-amplification coupled capture technique (EXACCT). In conclusion, the developed DNA isolation method can be used for the study of large numbers of stool samples, is independent of the age or method of stool collection and is suitable for large-scale screening studies.