Document Type

Journal Article

Department/Unit

School of Chinese Medicine

Title

Comparison of the methods for generating single-stranded DNA in SELEX

Language

English

Abstract

© 2015 The Royal Society of Chemistry.The generation of single-stranded DNA (ssDNA) from double-stranded PCR products is an essential step in the selection of aptamers by systematic evolution of ligands by exponential enrichment (SELEX). Magnetic separation with streptavidin-coated beads is always the most commonly used method. Recently, two size separation methods derived from unequal primers with chemical or structural modification were designed in SELEX. In this report, we made a comparison between magnetic separation and the two size separation methods for generation of ssDNA from double-stranded PCR products. Our results showed that all the methods produced ssDNA of good purity. Compared to the magnetic separation, size separation derived from unequal primers with chemical modification achieved an almost equivalent recovery rate of ssDNA, whereas size separation derived from unequal primers with structural modification showed a lower recovery rate of ssDNA. Considering the low cost, size separation derived from unequal primers with chemical modification could be a satisfactory alternative to the classic magnetic separation for the generation of ssDNA in SELEX.

Publication Date

2015

Source Publication Title

Analyst

Volume

140

Issue

10

Start Page

3439

End Page

3444

Publisher

Royal Society of Chemistry

DOI

10.1039/C5AN00244C

Link to Publisher's Edition

http://dx.doi.org/10.1039/C5AN00244C

ISSN (print)

00032654

ISSN (electronic)

13645528

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