An automated fluorescent single strand conformation polymorphism technique for high throughput mutation screening
(2001) In Chinese Medical Journal 114(11). p.1147-1150- Abstract
- 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%)... (More)
- 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. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1122517
- author
- Weng, Jianping ; Lehto, Markku ; Berglund, Anna and Groop, Leif LU
- organization
- publishing date
- 2001
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Chinese Medical Journal
- volume
- 114
- issue
- 11
- pages
- 1147 - 1150
- publisher
- Chinese Medical Association
- external identifiers
-
- pmid:11729508
- scopus:0035177561
- ISSN
- 0366-6999
- language
- English
- LU publication?
- yes
- id
- b2418db6-e348-4ce5-b2ea-c109c6e24226 (old id 1122517)
- alternative location
- http://www.cmj.org/Periodical/PDF/2001/2001111147.pdf
- date added to LUP
- 2016-04-01 16:07:08
- date last changed
- 2024-01-11 01:48:46
@article{b2418db6-e348-4ce5-b2ea-c109c6e24226, abstract = {{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.}}, author = {{Weng, Jianping and Lehto, Markku and Berglund, Anna and Groop, Leif}}, issn = {{0366-6999}}, language = {{eng}}, number = {{11}}, pages = {{1147--1150}}, publisher = {{Chinese Medical Association}}, series = {{Chinese Medical Journal}}, title = {{An automated fluorescent single strand conformation polymorphism technique for high throughput mutation screening}}, url = {{http://www.cmj.org/Periodical/PDF/2001/2001111147.pdf}}, volume = {{114}}, year = {{2001}}, }