Pyrosequencing protocol requiring a unique biotinylated primer
作者： Jose Luis Royo , Manuel Hidalgo Pascual , Ana Salinas , Francisco Jose Tello , Maria del Carmen Rivero
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摘要: Background: DNA sequencing has markedly changed the nature of biomedical research. Large-scale projects have generated several millions potential polymorphisms widespread in human genome requiring validation and incorporation into screening panels. As a consequence, high-throughput analysis these variants different populations interest is now cornerstone structural genomics. Pyrosequencing versatile technique allowing an easy 96-well typing format. However, every polymorphism requires specific labeled primer to generate single-stranded fragment containing region interest. Methods: We describe how with adjusted stoichiometry we can standardize labeling amplicon single biotinylated universal (BM13S). Results: circumvent need for primers each single-nucleotide (SNP) under study. example, assessed this novel protocol by genotyping three SNPs mapping calpain-10, caveolin-1 CYP19A1. Conclusion: The present approach represents alternative standard pyrosequencing protocols, since it that suitable SNP
sci-hub.se 参考文章(16)
M. Ronaghi, DNA SEQUENCING:A Sequencing Method Based on Real-Time Pyrophosphate Science. ,vol. 281, pp. 363- 365 ,(1998) , 10.1126/SCIENCE.281.5375.363
ULRICH H. FREY, HOLGER NÜCKEL, DOBROMIR DOBREV, IRIS MANTHEY, I. E. SANDALCIOGLU, ANDREAS EISENHARDT, WORM KARL, HAUNER HANS, WINFRIED SIFFERT, Quantification of G protein Gaalphas subunit splice variants in different human tissues and cells using pyrosequencing. Gene Expression. ,vol. 12, pp. 69- 81 ,(2005) , 10.3727/000000005783992124
Paul Hardenbol, Johan Banér, Maneesh Jain, Mats Nilsson, Eugeni A Namsaraev, George A Karlin-Neumann, Hossein Fakhrai-Rad, Mostafa Ronaghi, Thomas D Willis, Ulf Landegren, Ronald W Davis, Multiplexed genotyping with sequence-tagged molecular inversion probes Nature Biotechnology. ,vol. 21, pp. 673- 678 ,(2003) , 10.1038/NBT821
Hossein Fakhrai-Rad, Nader Pourmand, Mostafa Ronaghi, Pyrosequencing: An accurate detection platform for single nucleotide polymorphisms Human Mutation. ,vol. 19, pp. 479- 485 ,(2002) , 10.1002/HUMU.10078
Christina L. Aquilante, Maximilian T. Lobmeyer, Taimour Y. Langaee, Julie A. Johnson, Comparison of cytochrome P450 2C9 genotyping methods and implications for the clinical laboratory. Pharmacotherapy. ,vol. 24, pp. 720- 726 ,(2004) , 10.1592/PHCO.24.8.720.36074
Tommy Nordström, Karim Nourizad, Mostafa Ronaghi, Pål Nyrén, Method Enabling Pyrosequencing on Double-Stranded DNA Analytical Biochemistry. ,vol. 282, pp. 186- 193 ,(2000) , 10.1006/ABIO.2000.4603
Baback Gharizadeh, Jonas Eriksson, Nader Nourizad, Tommy Nordström, Pål Nyrén, Improvements in Pyrosequencing technology by employing Sequenase polymerase. Analytical Biochemistry. ,vol. 330, pp. 272- 280 ,(2004) , 10.1016/J.AB.2004.03.018
Erik Söderbäck, Anna-Lena Zackrisson, Bertil Lindblom, Anders Alderborn, Determination of CYP2D6 Gene Copy Number by Pyrosequencing Clinical Chemistry. ,vol. 51, pp. 522- 531 ,(2005) , 10.1373/CLINCHEM.2004.043182
Louise Nordfors, M�rten Jansson, Gunnar Sandberg, Catharina Lavebratt, Selim Sengul, Martin Schalling, Peter Arner, Large-scale genotyping of single nucleotide polymorphisms by Pyrosequencing™ and validation against the 5′nuclease (Taqman®) assay Human Mutation. ,vol. 19, pp. 395- 401 ,(2002) , 10.1002/HUMU.10062
Sanjay Tyagi, Diana P. Bratu, Fred Russell Kramer, Multicolor molecular beacons for allele discrimination Nature Biotechnology. ,vol. 16, pp. 49- 53 ,(1998) , 10.1038/NBT0198-49