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Development of an automated SNP analysis method using a paramagnetic beads handling robot
Author(s) -
Hagiwara Hiroko,
SawakamiKobayashi Kazumi,
Yamamoto Midori,
Iwasaki Shoji,
Sugiura Mika,
Abe Hatsumi,
KunihiroOhashi Sumiko,
Takase Kumiko,
Yamane Noriko,
Kato Kaoru,
Son Renkon,
Nakamura Michihiro,
Segawa Osamu,
Yoshida Mamiko,
Yohda Masafumi,
Tajima Hideji,
Kobori Masato,
Takahama Yousuke,
Itakura Mitsuo,
Machida Masayuki
Publication year - 2007
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.21380
Subject(s) - typing , snp , multiplex , adapter (computing) , single nucleotide polymorphism , computational biology , biology , genetics , genotype , computer science , gene , computer hardware
Abstract Biological and medical importance of the single nucleotide polymorphism (SNP) has led to development of a wide variety of methods for SNP typing. Aiming for establishing highly reliable and fully automated SNP typing, we have developed the adapter ligation method in combination with the paramagnetic beads handling technology, Magtration®. The method utilizes sequence specific ligation between the fluorescently labeled adapter and the sample DNAs at the cohesive end produced by a type IIS restriction enzyme. Evaluation of the method using human genomic DNA showed clear discrimination of the three genotypes without ambiguity using the same reaction condition for any SNPs examined. The operations following PCR amplification were automatically performed by the Magtration®‐based robot that we have previously developed. Multiplex typing of two SNPs in a single reaction by using four fluorescent dyes was successfully preformed at the almost same sensitivity and reliability as the single typing. These results demonstrate that the automated paramagnetic beads handling technology, Magtration®, is highly adaptable to the automated SNP analysis and that our method best fits to an automated in‐house SNP typing for laboratory and medical uses. Biotechnol. Bioeng. 2007;98; 420–428. © 2007 Wiley Periodicals, Inc.