IMPROVED SIZING OF FRAGILE-X CCG REPEATS BY NESTED POLYMERASE CHAIN-REACTION

被引：38

作者：

LEVINSON, G ^{[1
]}

MADDALENA, A ^{[1
]}

PALMER, FT ^{[1
]}

HARTON, GL ^{[1
]}

BICK, DP ^{[1
]}

HOWARDPEEBLES, PN ^{[1
]}

BLACK, SH ^{[1
]}

SCHULMAN, JD ^{[1
]}

机构：

[1] VIRGINIA COMMONWEALTH UNIV,MED COLL VIRGINIA,DEPT HUMAN GENET,RICHMOND,VA 23298

来源：

AMERICAN JOURNAL OF MEDICAL GENETICS | 1994年 / 51卷 / 04期

关键词：

MENTAL RETARDATION; GENETIC SCREENING; FMR-1; TRINUCLEOTIDE REPEATS;

D O I：

10.1002/ajmg.1320510448

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

We have developed an improved method for polymerase chain reaction (PCR)-based sizing of the CCG repeat region at the fragile X locus, FMR-1. This method is designed to optimize denaturation and replication of long repeats with high G + C content, which are otherwise refractory to amplification. The method utilizes nested PCR primers to increase sensitivity and specificity. Alkaline denaturation of the genomic template DNA, combined with addition of glycerol and deaza-dGTP, facilitates strand separation. Labeled PCR products are sized on denaturing polyacrylamide gels. For alleles in the normal-to-premutation size range, strong reproducible signals are routinely obtained from small amounts of rapidly prepared DNA. This allows precise determination of the CCG repeat number, providing data related to the expansion potential of the repetitive segment. Detection of large premutations and some full mutations is also enhanced by the improved procedure. (C) 1994 Wiley-Liss, Inc.

引用

页码：527 / 534

页数：8

共 21 条

[1]

CUSI MG, 1992, BIOTECHNIQUES, V12, P502

[2] RAPID CLONING OF HLA-A,B CDNA BY USING THE POLYMERASE CHAIN-REACTION - FREQUENCY AND NATURE OF ERRORS PRODUCED IN AMPLIFICATION [J].

ENNIS, PD ;

ZEMMOUR, J ;

SALTER, RD ;

PARHAM, P .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1990, 87 (07) :2833-2837

[3]

ERSTER SH, 1992, HUM GENET, V90, P55

[4]

FU YH, 1991, CELL, V67, P1

[5] A STUDY OF THE ORIGIN OF SHADOW BANDS SEEN WHEN TYPING DINUCLEOTIDE REPEAT POLYMORPHISMS BY THE PCR [J].

HAUGE, XY ;

LITT, M .

HUMAN MOLECULAR GENETICS, 1993, 2 (04) :411-415

[6] INHERITANCE OF THE FRAGILE-X SYNDROME - SIZE OF THE FRAGILE-X PREMUTATION IS A MAJOR DETERMINANT OF THE TRANSITION TO FULL MUTATION [J].

HEITZ, D ;

DEVYS, D ;

IMBERT, G ;

KRETZ, C ;

MANDEL, JL .

JOURNAL OF MEDICAL GENETICS, 1992, 29 (11) :794-801

[7] MOLECULAR GENETIC DIAGNOSIS OF SICKLE-CELL DISEASE USING DRIED BLOOD SPECIMENS ON BLOTTERS USED FOR NEWBORN SCREENING [J].

JINKS, DC ;

MINTER, M ;

TARVER, DA ;

VANDERFORD, M ;

HEJTMANCIK, JF ;

MCCABE, ERB .

HUMAN GENETICS, 1989, 81 (04) :363-366

[8] MAPPING OF DNA INSTABILITY AT THE FRAGILE-X TO A TRINUCLEOTIDE REPEAT SEQUENCE P(CCG)N [J].

KREMER, EJ ;

PRITCHARD, M ;

LYNCH, M ;

YU, S ;

HOLMAN, K ;

BAKER, E ;

WARREN, ST ;

SCHLESSINGER, D ;

SUTHERLAND, GR ;

RICHARDS, RI .

SCIENCE, 1991, 252 (5013) :1711-1714

[9]

Lemire M, 1993, Union Med Can, V122, P23

[10] RELIABLE GENDER SCREENING FOR HUMAN PREIMPLANTATION EMBRYOS, USING MULTIPLE DNA TARGET-SEQUENCES [J].

LEVINSON, G ;

FIELDS, RA ;

HARTON, GL ;

PALMER, FT ;

MADDALENA, A ;

FUGGER, EF ;

SCHULMAN, JD .

HUMAN REPRODUCTION, 1992, 7 (09) :1304-1313

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