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Leif Groop

Leif Groop

Principal investigator

Leif Groop

An automated fluorescent single strand conformation polymorphism technique for high throughput mutation screening

Author

  • Jianping Weng
  • Markku Lehto
  • Anna Berglund
  • Leif Groop

Summary, in English

OBJECTIVE: To develop a high throughput mutational detection method by multiple fluorescence-labeled polymerase chain reaction (PCR) products. METHODS: A total of 27 known mutations including 22 substitutions, 3 insertions (1, 2 and 7 bp) and 2 deletions (1 and 2 bp) in the hepatocyte nuclear factor (HNF)-4 alpha, glucokinase and HNF-1 alpha genes were tested. During nested PCR, amplified fragments were labeled with three fluorescent dyes. PCR products were visualized with an ABI-377 fluorescence sequencer using 5% glycerol or 10% sucrose in non-denaturing gel conditions. RESULTS: Twenty-five of 27 variants (93%) could be detected by combining 5% glycerol and 10% sucrose gel matrix conditions. Twenty-two of 27 (82%) and 18 of 27 (67%) variants were identified using 5% glycerol and 10% sucrose conditions, respectively. CONCLUSION: This fluorescence-based PCR single strand conformation polymorphism technique represents a simple, non-hazardous, time-saving and sensitive method for high throughput mutation detection.

Department/s

  • Genomics, Diabetes and Endocrinology

Publishing year

2001

Language

English

Pages

1147-1150

Publication/Series

Chinese Medical Journal

Volume

114

Issue

11

Document type

Journal article

Publisher

Chinese Medical Association

Topic

  • Endocrinology and Diabetes

Status

Published

Research group

  • Genomics, Diabetes and Endocrinology

ISBN/ISSN/Other

  • ISSN: 0366-6999