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

被引：29

作者：

Gennarelli, M

Pavoni, M

Amicucci, P

Novelli, G

Dallapiccola, B

机构：

[1] Univ Roma Tor Vergata, Cattedra Genet Umana, Dipartimento Biopatol & Diagnost Immagini, Fac Med & Chirurg,Chair Human & Med Genet, I-00133 Rome, Italy

[2] CSS Mendel Inst, Rome, Italy

来源：

DIAGNOSTIC MOLECULAR PATHOLOGY | 1998年 / 7卷 / 03期

关键词：

long PCR; myotonic dystrophy; CTG repeats;

D O I：

10.1097/00019606-199806000-00002

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 (greater than or equal to 15 pg). This method is quick, sensitive, and reproducible and reduces the cost of diagnostic laboratory processing.

引用

页码：135 / 137

页数：3

共 14 条

[1] MOLECULAR-BASIS OF MYOTONIC-DYSTROPHY - EXPANSION OF A TRINUCLEOTIDE (CTG) REPEAT AT THE 3' END OF A TRANSCRIPT ENCODING A PROTEIN-KINASE FAMILY MEMBER [J].

BROOK, JD ;

MCCURRACH, ME ;

HARLEY, HG ;

BUCKLER, AJ ;

CHURCH, D ;

ABURATANI, H ;

HUNTER, K ;

STANTON, VP ;

THIRION, JP ;

HUDSON, T ;

SOHN, R ;

ZEMELMAN, B ;

SNELL, RG ;

RUNDLE, SA ;

CROW, S ;

DAVIES, J ;

SHELBOURNE, P ;

BUXTON, J ;

JONES, C ;

JUVONEN, V ;

JOHNSON, K ;

HARPER, PS ;

SHAW, DJ ;

HOUSMAN, DE .

CELL, 1992, 68 (04) :799-808

[2]

Cheng S, 1996, HUM MUTAT, V7, P304, DOI 10.1002/(SICI)1098-1004(1996)7:4<304::AID-HUMU3>3.3.CO

[3]

2-4

[4] AN UNSTABLE TRIPLET REPEAT IN A GENE RELATED TO MYOTONIC MUSCULAR-DYSTROPHY [J].

FU, YH ;

PIZZUTI, A ;

FENWICK, RG ;

KING, J ;

RAJNARAYAN, S ;

DUNNE, PW ;

DUBEL, J ;

NASSER, GA ;

ASHIZAWA, T ;

DEJONG, P ;

WIERINGA, B ;

KORNELUK, R ;

PERRYMAN, MB ;

EPSTEIN, HF ;

CASKEY, CT .

SCIENCE, 1992, 255 (5049) :1256-1258

[5]

Gennarelli M, 1996, AM J MED GENET, V65, P342, DOI 10.1002/(SICI)1096-8628(19961111)65:4<342::AID-AJMG18>3.0.CO

[6]

2-U

[7]

GUIDA M, 1995, CLIN CHEM, V41, P69

[8]

HARPER PS, 1989, MYOTONIC DYSTROPHY, P316

[9]

Hecimovic S, 1997, CLIN GENET, V52, P147

[10]

JANSEN G, 1994, AM J HUM GENET, V54, P575

← 1 2 →