Determination of copy number of short tandem repeat using NAD-dependent ligase and pyrosequencing-compatible method(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)

スポンサーリンク

概要

• 論文の詳細を見る

• A sensitive and pyrosequencing-compatible method for determining the copy number of the short tandem repeat (STR) is presented in this study. When Escherichia coli ligase catalyzes the ligation of primer and probes complementary to the proper sites of the target DNA template, it converts nicotinamide adenine dinucleotide to adenosine mono-phosphate (AMP) and nicotinamide. The AMP release level is proportional to the copy number of the STR and can be measured using adenylate kinase, pyruvate kinase, and luciferase. Unlike current standard methods based on electrophoresis, the present assay is sensitive to the point mutation. Furthermore, after determination of the copy number of the tandem repeat using the proposed method, the DNA templates, primer and probes immobilized onto super paramagnetic beads can be washed and pyrosequencing can be applied for the remaining DNA sequencing. This assay is specially efficient to handle a large number of samples because massively parallel tests could be executed in a microplate photometer. Furthermore, it can work with the pyrosequencing for further sequencing like genome sequencing.

• 公益社団法人日本生物工学会の論文

著者

• Lin Ming-Shu

  Department of Chemical Engineering, National Taipei University of Technology:Institute of Biotechnology, National Taipei University of Technology

• Yao Ciou-Hao

  Department of Chemical Engineering, National Taipei University of Technology

• Lu Cheng-Hung

  Institute of Biotechnology, National Taipei University of Technology

• Hou Shao-Yi

  Department of Chemical Engineering, National Taipei University of Technology

• Hou Shao-Yi

  Department of Chemical Engineering, National Taipei University of Technology:Institute of Biotechnology, National Taipei University of Technology

関連論文

• Determination of copy number of short tandem repeat using NAD-dependent ligase and pyrosequencing-compatible method(GENETICS, MOLECULAR BIOLOGY, AND GENE ENGINEERING)

スポンサーリンク