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Determination of copy number of short tandem repeat using NAD-dependent ligase and pyrosequencing-compatible method

Paper ID Volume ID Publish Year Pages File Format Full-Text
20549 43180 2013 5 PDF Available
Title
Determination of copy number of short tandem repeat using NAD-dependent ligase and pyrosequencing-compatible method
Abstract

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 monophosphate (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.

Keywords
Short tandem repeat; NAD-dependent ligase; Pyrosequencing; Pyruvate kinase; Adenylate kinase
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Determination of copy number of short tandem repeat using NAD-dependent ligase and pyrosequencing-compatible method
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 116, Issue 5, November 2013, Pages 546–550
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us