A Multiplex Ligation Assay for miRNA Copy Number Profiling

Duncan Kilburn; Yunke Song; Tza-Huei Wang; Kelvin J Liu

doi:10.1007/978-1-4939-6524-3_17

A Multiplex Ligation Assay for miRNA Copy Number Profiling

Methods Mol Biol. 2017:1509:185-193. doi: 10.1007/978-1-4939-6524-3_17.

Authors

Duncan Kilburn¹, Yunke Song², Tza-Huei Wang^{2

3

4

5}, Kelvin J Liu⁶

Affiliations

¹ Circulomics Inc., 810 Wyman Park Drive, Unit G01, Baltimore, MD, 21211, USA. [email protected].
² Biomedical Engineering Department, Johns Hopkins University, Baltimore, MD, USA.
³ Mechanical Engineering Department, Johns Hopkins University, Baltimore, MD, USA.
⁴ Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD, USA.
⁵ Center of Cancer Nanotechnology Excellence, Johns Hopkins University, Baltimore, MD, USA.
⁶ Circulomics Inc., 810 Wyman Park Drive, Unit G01, Baltimore, MD, 21211, USA.

PMID: 27826928
DOI: 10.1007/978-1-4939-6524-3_17

Abstract

Ligo-miR is an assay technology that can perform multiplexed detection of miRNAs from a wide range of biological sources. At its core are two sequential ligation steps. First in the capture ligation, template molecules are created by ligating a DNA adapter to the 3' end of all miRNA molecules. Then in the coding ligation these templates are used to generate, linearly amplified, DNA products encoded by length. The resultant number of each DNA product is proportional to the original number of miRNA molecules. The products and their corresponding miRNA can be identified and quantified using common DNA sizing methods such as electrophoresis.

Keywords: Copy number; Ligation; Multiplex; Profiling; Quantification assay; miRNA.

MeSH terms

Cells, Cultured
Gene Expression Profiling / methods*
Humans
MicroRNAs / genetics*
MicroRNAs / isolation & purification
MicroRNAs / metabolism

Substances

MicroRNAs

Grants and funding

R43 GM103360/GM/NIGMS NIH HHS/United States