DNA extraction from archived specimens by sonication

Robert A. Robinson, Michael (Mike) Heller

Research output: Contribution to journalArticle

Abstract

The analysis of DNA from archived tissues is an important clinical research and diagnostic tool. The analyses of these preserved tissues that are most commonly formalin-fixed and paraffin-embedded allow for the retrospective study of a variety of genetic changes, or even the presence of infectious agents. Prior to polymerase chain reaction (PCR), the genetic analyses of fixed tissues were carried out using in situ hybridization, dot-blot hybridization, or Southern blotting. These methodologies have traditionally provided some degree of success, but are sometimes limited in sensitivity, involve complex procedures, or cannot detect degraded DNA, which is found in old-fixed tissues. PCR analysis provides a simple and sensitive method for the analysis of DNA from fixed tissues. A number of methods exist for isolating DNA from formalin-fixed, paraffin-embedded tissues, but the best procedures in terms of reliably releasing DNA involve a large number of solvent extractions, centrifugations, and long digestion periods with enzyme buffers. A method has been developed that quickly and efficiently extracts DNA from paraffin-embedded tissue. This method utilizes a sonicating water bath to disrupt the tissue samples and such samples can be prepared in less than 30 min with only solvent extraction in a minimum of steps.

Original languageEnglish (US)
Pages (from-to)431-443
Number of pages13
JournalMethods in Neurosciences
Volume26
Issue numberC
DOIs
StatePublished - Jan 1 1995
Externally publishedYes

Fingerprint

Sonication
DNA
Paraffin
Formaldehyde
Polymerase Chain Reaction
Southern Blotting
Centrifugation
Baths
In Situ Hybridization
Digestion
Buffers
Retrospective Studies
Water
Enzymes
Research

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

DNA extraction from archived specimens by sonication. / Robinson, Robert A.; Heller, Michael (Mike).

In: Methods in Neurosciences, Vol. 26, No. C, 01.01.1995, p. 431-443.

Research output: Contribution to journalArticle

@article{2f944bfe2a3e4644ac8157d26200dcc7,
title = "DNA extraction from archived specimens by sonication",
abstract = "The analysis of DNA from archived tissues is an important clinical research and diagnostic tool. The analyses of these preserved tissues that are most commonly formalin-fixed and paraffin-embedded allow for the retrospective study of a variety of genetic changes, or even the presence of infectious agents. Prior to polymerase chain reaction (PCR), the genetic analyses of fixed tissues were carried out using in situ hybridization, dot-blot hybridization, or Southern blotting. These methodologies have traditionally provided some degree of success, but are sometimes limited in sensitivity, involve complex procedures, or cannot detect degraded DNA, which is found in old-fixed tissues. PCR analysis provides a simple and sensitive method for the analysis of DNA from fixed tissues. A number of methods exist for isolating DNA from formalin-fixed, paraffin-embedded tissues, but the best procedures in terms of reliably releasing DNA involve a large number of solvent extractions, centrifugations, and long digestion periods with enzyme buffers. A method has been developed that quickly and efficiently extracts DNA from paraffin-embedded tissue. This method utilizes a sonicating water bath to disrupt the tissue samples and such samples can be prepared in less than 30 min with only solvent extraction in a minimum of steps.",
author = "Robinson, {Robert A.} and Heller, {Michael (Mike)}",
year = "1995",
month = "1",
day = "1",
doi = "10.1016/S1043-9471(06)80106-0",
language = "English (US)",
volume = "26",
pages = "431--443",
journal = "Methods in Neurosciences",
issn = "1043-9471",
publisher = "Academic Press Inc.",
number = "C",

}

TY - JOUR

T1 - DNA extraction from archived specimens by sonication

AU - Robinson, Robert A.

AU - Heller, Michael (Mike)

PY - 1995/1/1

Y1 - 1995/1/1

N2 - The analysis of DNA from archived tissues is an important clinical research and diagnostic tool. The analyses of these preserved tissues that are most commonly formalin-fixed and paraffin-embedded allow for the retrospective study of a variety of genetic changes, or even the presence of infectious agents. Prior to polymerase chain reaction (PCR), the genetic analyses of fixed tissues were carried out using in situ hybridization, dot-blot hybridization, or Southern blotting. These methodologies have traditionally provided some degree of success, but are sometimes limited in sensitivity, involve complex procedures, or cannot detect degraded DNA, which is found in old-fixed tissues. PCR analysis provides a simple and sensitive method for the analysis of DNA from fixed tissues. A number of methods exist for isolating DNA from formalin-fixed, paraffin-embedded tissues, but the best procedures in terms of reliably releasing DNA involve a large number of solvent extractions, centrifugations, and long digestion periods with enzyme buffers. A method has been developed that quickly and efficiently extracts DNA from paraffin-embedded tissue. This method utilizes a sonicating water bath to disrupt the tissue samples and such samples can be prepared in less than 30 min with only solvent extraction in a minimum of steps.

AB - The analysis of DNA from archived tissues is an important clinical research and diagnostic tool. The analyses of these preserved tissues that are most commonly formalin-fixed and paraffin-embedded allow for the retrospective study of a variety of genetic changes, or even the presence of infectious agents. Prior to polymerase chain reaction (PCR), the genetic analyses of fixed tissues were carried out using in situ hybridization, dot-blot hybridization, or Southern blotting. These methodologies have traditionally provided some degree of success, but are sometimes limited in sensitivity, involve complex procedures, or cannot detect degraded DNA, which is found in old-fixed tissues. PCR analysis provides a simple and sensitive method for the analysis of DNA from fixed tissues. A number of methods exist for isolating DNA from formalin-fixed, paraffin-embedded tissues, but the best procedures in terms of reliably releasing DNA involve a large number of solvent extractions, centrifugations, and long digestion periods with enzyme buffers. A method has been developed that quickly and efficiently extracts DNA from paraffin-embedded tissue. This method utilizes a sonicating water bath to disrupt the tissue samples and such samples can be prepared in less than 30 min with only solvent extraction in a minimum of steps.

UR - http://www.scopus.com/inward/record.url?scp=77957085003&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=77957085003&partnerID=8YFLogxK

U2 - 10.1016/S1043-9471(06)80106-0

DO - 10.1016/S1043-9471(06)80106-0

M3 - Article

VL - 26

SP - 431

EP - 443

JO - Methods in Neurosciences

JF - Methods in Neurosciences

SN - 1043-9471

IS - C

ER -