Whole Genome Amplification from Circulating Tumor Cells
A ligation-adaptor method of WGA was performed on whole cell lysates from pooled CTCs isolated by IE/FACS using a GenomePlex whole genome amplification kit (WGA4, Sigma-Aldrich) according to the manufacturer’s instructions [12 (link),25 (link)]. DNA was randomly fragmented and converted to polymerase chain reaction (PCR)-amplifiable library molecules flanked by universal priming sites. PCR amplification of library molecules was performed using universal oligonucleotide primers.
Kelley R.K., Magbanua M.J., Butler T.M., Collisson E.A., Hwang J., Sidiropoulos N., Evason K., McWhirter R.M., Hameed B., Wayne E.M., Yao F.Y., Venook A.P, & Park J.W. (2015). Circulating tumor cells in hepatocellular carcinoma: a pilot study of detection, enumeration, and next-generation sequencing in cases and controls. BMC Cancer, 15, 206.
Other organizations :
University of California, San Francisco, UCSF Helen Diller Family Comprehensive Cancer Center, Oregon Health & Science University, University of Vermont Medical Center
DNA randomly fragmented and converted to PCR-amplifiable library molecules
control variables
Manufacturer's instructions for GenomePlex whole genome amplification kit (WGA4, Sigma-Aldrich)
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
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