𝔖 Bobbio Scriptorium
✦   LIBER   ✦

Detection of single nucleotide polymorphisms in coagulation factor XI deficient patients by multitemperature single-strand conformation polymorphism analysis

✍ Scribed by Alexandra Bezak; Radosław Kaczanowski; Astrid Dossenbach-Glaninger; Krzysztof Kucharczyk; Werner Lubitz; Pierre Hopmeier

Publisher
John Wiley and Sons
Year
2005
Tongue
English
Weight
242 KB
Volume
19
Category
Article
ISSN
0887-8013

No coin nor oath required. For personal study only.

✦ Synopsis

Factor XI (FXI) deficiency is a rare inherited disorder which can cause bleeding complications especially in case of hemostatic challenge and/or in tissues with high fibrinolytic activity. A number of causative mutations have been described in FXI deficient individuals which have been detected by various screening methods. In this study, we present the application of the multitemperature single-strand conformation polymorphism analysis (MSSCP) on the FXI gene, a recently developed methodology for the detection of single nucleotide exchanges. We analyzed a total of 217 polymerase chain reaction (PCR) fragments from the promoter region as well as from exons 1-7 and 11-15 and compared the results to automatic fluorescent sequencing. A total of 29 PCR fragments showed single nucleotide exchanges in conventional fluorescent sequencing, representing 10 different mutations (nine missense mutations, one small deletion) and four frequent polymorphisms. With MSSCP electrophoresis at a standard temperature profile (gel temperature 35-20-101C) we were able to detect 13 of 14 (93%) different nucleotide exchanges in 25 of 29 PCR fragments (86%). Hence, the detection rate for genetic variations in the FXI gene was 86%. To evaluate the reproducibility, MSSCP was performed twice for 174 PCR fragments and the consistency between two electrophoretic runs was 99%. We conclude that the MSSCP is a sensitive, fast, and cost effective screening method for the detection of FXI gene mutations.

📜 SIMILAR VOLUMES

Detection of point mutations in the chlo
Detection of point mutations in the chloroplast genome by single-stranded conformation polymorphism analysis
✍ Kin-Ying To; Chiu-I. Liu; Shih-Tung Liu; Yu-Sun Chang 📂 Article 📅 1993 🏛 John Wiley and Sons 🌐 English ⚖ 465 KB

## Summary Single‐stranded conformation polymorphism (SSCP) analysis was used to examine the mutations of the chloroplast 16S rRNA locus of streptomycin‐resistant mutants in __Nicotiana plumbaginifolia.__ DNA fragments of 121, 517, 968 and 1578 bp, each possessing a known point mutation, were gener

Detection of mutations and polymorphisms
Detection of mutations and polymorphisms in the p53 tumor suppressor gene by single-strand conformation polymorphism analysis
✍ Laura S. Kutach; Svetlana Bolshakov; Honnavara N. Ananthaswamy 📂 Article 📅 1999 🏛 John Wiley and Sons 🌐 English ⚖ 124 KB 👁 2 views

## Detection of mutations and polymorphisms in the p53 tumor suppressor gene by single-strand conformation polymorphism analysis Deciphering the genetic mechanisms in cancer development requires analysis of a large number of tumors for consistent genetic alterations. Single-strand conformational p

Detection of known and new mutations in
Detection of known and new mutations in the thiopurine S-methyltransferase gene by single-strand conformation polymorphism analysis
✍ Catherine Spire-Vayron de la Moureyre; Hervé Debuysère; Nadda Sabbagh; Delphine 📂 Article 📅 1998 🏛 John Wiley and Sons 🌐 English ⚖ 467 KB 👁 2 views

To detect mutations in the thiopurine S-methyltransferase gene (TPMT), we have developed a strategy based on single-strand conformation polymorphism (SSCP) analysis of the gene amplified by polymerase chain reaction (PCR). The sensitivity of the method was first evaluated by analyzing DNA samples fr