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Nucleic Acids Research
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Nucleic Acids Research
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microDuMIP: target-enrichment technique for microarray-based duplex molecular inversion probes

Authors: Soo Min Han; Min Goo Lee; Duhee Bang; Jung Ki Yoon; Hoon Jang; Jinwoo Ahn; Ji Hyun Lee; +1 Authors

microDuMIP: target-enrichment technique for microarray-based duplex molecular inversion probes

Abstract

Molecular inversion probe (MIP)-based capture is a scalable and effective target-enrichment technology that can use synthetic single-stranded oligonucleotides as probes. Unlike the straightforward use of synthetic oligonucleotides for low-throughput target capture, high-throughput MIP capture has required laborious protocols to generate thousands of single-stranded probes from DNA microarray because of multiple enzymatic steps, gel purifications and extensive PCR amplifications. Here, we developed a simple and efficient microarray-based MIP preparation protocol using only one enzyme with double-stranded probes and improved target capture yields by designing probes with overlapping targets and unique barcodes. To test our strategy, we produced 11 510 microarray-based duplex MIPs (microDuMIPs) and captured 3554 exons of 228 genes in a HapMap genomic DNA sample (NA12878). Under our protocol, capture performance and precision of calling were compatible to conventional MIP capture methods, yet overlapping targets and unique barcodes allowed us to precisely genotype with as little as 50 ng of input genomic DNA without library preparation. microDuMIP method is simpler and cheaper, allowing broader applications and accurate target sequencing with a scalable number of targets.

Related Organizations
Keywords

Exome/genetics, DNA, Single-Stranded, Molecular Probe Techniques, Molecular Probe Techniques*, Polymerase Chain Reaction, Polymorphism, Single Nucleotide, Oligonucleotide Probes/genetics*, Exome, Polymorphism, Oligonucleotide Array Sequence Analysis, Reproducibility of Results, DNA, Single Nucleotide, Sequence Analysis, DNA, 543, DNA/methods, Oligonucleotide Array Sequence Analysis/methods*, Methods Online, Single-Stranded/genetics*, Oligonucleotide Probes, Sequence Analysis

  • BIP!
    Impact byBIP!
    citations
    This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    12
    popularity
    This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
    Average
    influence
    This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
    Average
    impulse
    This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
    Average
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citations
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
12
Average
Average
Average
Green
gold