A single polymerase chain reaction-based protocol for detecting normal and expanded alleles in myotonic dystrophy

Massimo Gennarelli, Marco Pavoni, Paola Amicucci, Giuseppe Novelli, Bruno Dallapiccola

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

The myotonic dystrophy (DM) expansion varies from 50 to 4000 CTG repeats in the 3' untranslated region of the DMPK gene. Direct analysis by Southern blot, after restriction enzyme digestion of genomic DNA, is the method of choice for studying the DM mutation. A long polymerase chain reaction (PCR)formatted protocol, which involved a single genomic in vitro amplification followed by high concentration agarose gel electrophoresis and oligo-specific hybridization, was used to amplify normal alleles and DM alleles in all examined ranges of expansion (up to 3,700 CTGs) starting from a small amount of genomic DNA (≥15 pg). This method is quick, sensitive, and reproducible and reduces the cost of diagnostic laboratory processing.

Original languageEnglish
Pages (from-to)135-137
Number of pages3
JournalDiagnostic Molecular Pathology
Volume7
Issue number3
DOIs
Publication statusPublished - 1998

Fingerprint

Myotonic Dystrophy
Alleles
Polymerase Chain Reaction
Agar Gel Electrophoresis
DNA
3' Untranslated Regions
Southern Blotting
Digestion
Costs and Cost Analysis
Mutation
Enzymes
Genes
In Vitro Techniques

Keywords

  • CTG repeats
  • Long PCR
  • Myotonic dystrophy

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

A single polymerase chain reaction-based protocol for detecting normal and expanded alleles in myotonic dystrophy. / Gennarelli, Massimo; Pavoni, Marco; Amicucci, Paola; Novelli, Giuseppe; Dallapiccola, Bruno.

In: Diagnostic Molecular Pathology, Vol. 7, No. 3, 1998, p. 135-137.

Research output: Contribution to journalArticle

@article{22207eca4f444dea8b854ff085a45535,
title = "A single polymerase chain reaction-based protocol for detecting normal and expanded alleles in myotonic dystrophy",
abstract = "The myotonic dystrophy (DM) expansion varies from 50 to 4000 CTG repeats in the 3' untranslated region of the DMPK gene. Direct analysis by Southern blot, after restriction enzyme digestion of genomic DNA, is the method of choice for studying the DM mutation. A long polymerase chain reaction (PCR)formatted protocol, which involved a single genomic in vitro amplification followed by high concentration agarose gel electrophoresis and oligo-specific hybridization, was used to amplify normal alleles and DM alleles in all examined ranges of expansion (up to 3,700 CTGs) starting from a small amount of genomic DNA (≥15 pg). This method is quick, sensitive, and reproducible and reduces the cost of diagnostic laboratory processing.",
keywords = "CTG repeats, Long PCR, Myotonic dystrophy",
author = "Massimo Gennarelli and Marco Pavoni and Paola Amicucci and Giuseppe Novelli and Bruno Dallapiccola",
year = "1998",
doi = "10.1097/00019606-199806000-00002",
language = "English",
volume = "7",
pages = "135--137",
journal = "Diagnostic Molecular Pathology",
issn = "1052-9551",
publisher = "Lippincott Williams and Wilkins",
number = "3",

}

TY - JOUR

T1 - A single polymerase chain reaction-based protocol for detecting normal and expanded alleles in myotonic dystrophy

AU - Gennarelli, Massimo

AU - Pavoni, Marco

AU - Amicucci, Paola

AU - Novelli, Giuseppe

AU - Dallapiccola, Bruno

PY - 1998

Y1 - 1998

N2 - The myotonic dystrophy (DM) expansion varies from 50 to 4000 CTG repeats in the 3' untranslated region of the DMPK gene. Direct analysis by Southern blot, after restriction enzyme digestion of genomic DNA, is the method of choice for studying the DM mutation. A long polymerase chain reaction (PCR)formatted protocol, which involved a single genomic in vitro amplification followed by high concentration agarose gel electrophoresis and oligo-specific hybridization, was used to amplify normal alleles and DM alleles in all examined ranges of expansion (up to 3,700 CTGs) starting from a small amount of genomic DNA (≥15 pg). This method is quick, sensitive, and reproducible and reduces the cost of diagnostic laboratory processing.

AB - The myotonic dystrophy (DM) expansion varies from 50 to 4000 CTG repeats in the 3' untranslated region of the DMPK gene. Direct analysis by Southern blot, after restriction enzyme digestion of genomic DNA, is the method of choice for studying the DM mutation. A long polymerase chain reaction (PCR)formatted protocol, which involved a single genomic in vitro amplification followed by high concentration agarose gel electrophoresis and oligo-specific hybridization, was used to amplify normal alleles and DM alleles in all examined ranges of expansion (up to 3,700 CTGs) starting from a small amount of genomic DNA (≥15 pg). This method is quick, sensitive, and reproducible and reduces the cost of diagnostic laboratory processing.

KW - CTG repeats

KW - Long PCR

KW - Myotonic dystrophy

UR - http://www.scopus.com/inward/record.url?scp=0031783721&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031783721&partnerID=8YFLogxK

U2 - 10.1097/00019606-199806000-00002

DO - 10.1097/00019606-199806000-00002

M3 - Article

C2 - 9836067

AN - SCOPUS:0031783721

VL - 7

SP - 135

EP - 137

JO - Diagnostic Molecular Pathology

JF - Diagnostic Molecular Pathology

SN - 1052-9551

IS - 3

ER -