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Chapter 7 : Nucleic Acid Isolation: Overview of Sample Preparation Methods

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Abstract:

The type of sample being submitted for analysis determines the method used for nucleic acid isolation. Serum is also an acceptable material and yields similar clinical utility, but viral quantification may be slightly lower than plasma from the same patient due to entrapment of some extracellular virus within the blood clot. Regardless of the source, nucleic acid extraction usually consists of three primary processes: (i) lysis, (ii) denaturation/degradation of other biomolecules, and (iii) separation of the nucleic acids from other constituents in the sample and/or concentration of the DNA or RNA. Although there are several possible ways to classify nucleic acid extraction techniques, the following types of chemistries are the most commonly used: (i) precipitative methods, (ii) liquid-phase extractions, and (iii) solid-phase extractions. Although all three types of extraction methods are used, solid-phase extractions now are the most widely utilized due to easy scalability and the availability of automated instruments for these methods. While precipitative and liquid-phase extraction methods are still widely used, the most commonly performed DNA and RNA extractions involve selective binding of nucleic acids to an immobilized matrix (solid phase). The majority of the alcohol can be removed by essentially drying the matrix with the nucleic acid bound. Automation of the nucleic acid extraction process typically provides a more reproducible yield of DNA and/or RNA.

Citation: Hill C. 2011. Nucleic Acid Isolation: Overview of Sample Preparation Methods, p 119-125. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch7

Key Concept Ranking

Bacterial Cell Wall
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Chromosomal DNA
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Nucleic Acids
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Sodium Dodecyl Sulfate
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Tables

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TABLE 1

Widely available automated nucleic acid extraction instruments

Citation: Hill C. 2011. Nucleic Acid Isolation: Overview of Sample Preparation Methods, p 119-125. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch7

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