Abstract
Droplet digital PCR (ddPCR) allows for accurate quantification of genetic events such as copy number variation and single nucleotide variants. Probe-based assays represent the current “gold-standard” for detection and quantification of these genetic events. Here, we introduce a cost-effective single color ddPCR assay that allows for single genome resolution quantification of copy number and single nucleotide variation.
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References
Gray ES, Rizos H, Reid AL, Boyd SC, Pereira MR, Lo J et al (2015) Circulating tumor DNA to monitor treatment response and detect acquired resistance in patients with metastatic melanoma. Oncotarget 6(39):42008–42018. https://doi.org/10.18632/oncotarget.5788
Miotke L, Lau BT, Rumma RT, Ji HP (2014) High sensitivity detection and quantitation of DNA copy number and single nucleotide variants with single color droplet digital PCR. Anal Chem 86(5):2618–2624. https://doi.org/10.1021/ac403843j
McDermott GP, Do D, Litterst CM, Maar D, Hindson CM, Steenblock ER et al (2013) Multiplexed target detection using DNA-binding dye chemistry in droplet digital PCR. Anal Chem 85(23):11619–11627. https://doi.org/10.1021/ac403061n
Rodriguez A, Laio A (2014) Machine learning. Clustering by fast search and find of density peaks. Science 344(6191):1492–1496. https://doi.org/10.1126/science.1242072
Acknowledgments
This work was supported by the National Institutes of Health (NIH) grant NHGR1 P01 HG00020526 (H.P.J., C.M.W.B).
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Wood-Bouwens, C.M., Ji, H.P. (2018). Single Color Multiplexed ddPCR Copy Number Measurements and Single Nucleotide Variant Genotyping. In: Karlin-Neumann, G., Bizouarn, F. (eds) Digital PCR. Methods in Molecular Biology, vol 1768. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7778-9_18
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DOI: https://doi.org/10.1007/978-1-4939-7778-9_18
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Publisher Name: Humana Press, New York, NY
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Online ISBN: 978-1-4939-7778-9
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