MUTATIONAL ANALYSIS USING DENATURING GRADIENT GEL-ELECTROPHORESIS AND PCR

被引：20

作者：

CARIELLO, NF

SKOPEK, TR

机构：

[1] University of North Carolina, Pathology, Department, Chapel Hill, NC

[2] University of North Carolina, Environmental Sciences and Engineering, Chapel Hill, NC

来源：

MUTATION RESEARCH | 1993年 / 288卷 / 01期

关键词：

DENATURING GRADIENT GEL ELECTROPHORESIS; PCR; DGGE; DNA MOLECULES; SEPARATION; SEQUENCE OF DNA MOLECULES; SEPARATION BASED ON; POLYMERASE CHAIN REACTION;

D O I：

10.1016/0027-5107(93)90212-X

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

Denaturing gradient gel electrophoresis (DGGE) separates (DNA) molecules based on their sequence. Using the proper conditions, all base-pair substitutions can be resolved from the wild-type sequence using DGGE. Polymerase chain reaction (PCR) permits rapid amplification of a given region of the genome. In this paper, we demonstrate the utility of DGGE combined with PCR for mutation analysis by presenting different examples: (i) analysis of mouse p53 cDNA for mutations, (ii) simultaneous analysis of thousands of 4NQO-induced mutants for mutations in HPRT exon 3, (iii) examination of the fidelity of the thermostable DNA polymerase isolated from Pyrococcus furiosus (Pfu), (iv) purification of mutant DNA from contaminating wild-type DNA from mouse spleenic T-cell clones.

引用

页码：103 / 112

页数：10

共 37 条

[1]

ATREE O, 1989, GENOMICS, V4, P266

[2] CONSTANT DENATURANT GEL-ELECTROPHORESIS AS A RAPID SCREENING TECHNIQUE FOR P53 MUTATIONS [J].