Measurement of copy number variation in single cancer cells using rapid-emulsification digital droplet MDA

作     者：Samuel C.Kim Gayatri Premasekharan Iain C.Clark Hawi B.Gemeda Pamela L.Paris Adam R.Abate 

作者机构：Department of Bioengineering and Therapeutic SciencesUniversity of CaliforniaSan Francisco(UCSF)California Institute for Quantitative Biosciences(QB3)San FranciscoSan FranciscoCA 94158USA Department of UrologyDivision of Hematology&OncologyUniversity of CaliforniaSan Francisco(UCSF)San FranciscoCA 94158USA 

出 版 物：《Microsystems & Nanoengineering》 (微系统与纳米工程（英文）)

年 卷 期：2017年第3卷第1期

页      面：260-266页

核心收录：

学科分类：1002[医学-临床医学] 100214[医学-肿瘤学] 10[医学] 

基　　金：We thank Angus Sidore and Freeman Lan for helpful scientific discussions.This work was supported by the UCSF Division of Hematology-Oncology Perkins Philanthropy(PLP) the National Science Foundation CAREER Award(Grant Number DBI-1253293) the National Institutes of Health(NIH)(Grant Numbers HG007233-01,R01-EB019453-01,and DP2-AR068129-01) the Defense Advanced Research Projects Agency Living Foundries Program(Contract Numbers HR0011-12-C-0065,N66001-12-C-4211,and HR0011-12-C-0066) Fold F(x)Program(Contract Number DE-AC02-05CH11231) 

主　　题：amplification bias copy number variation ddMDA droplet microfluidics multiple displacement amplification 

摘      要：Uniform amplification of low-input DNA is important for applications across biology,including single-cell genomics,forensic science,and microbial and viral ***,the requisite biochemical amplification methods are prone to bias,skewing sequence proportions and obscuring signals relating to copy *** droplet multiple displacement amplification enables uniform amplification but requires expert knowledge of microfluidics to generate monodisperse *** addition,existing microfluidic methods are tedious and labor intensive for preparing many ***,we introduce rapid-emulsification multiple displacement amplification,a method to generate monodisperse droplets with a hand-held syringe and hierarchical droplet *** conventional microfluidic devices require 10 min to emulsify a sample,our system requires tens of seconds and yields data of equivalent *** demonstrate the approach by using it to accurately measure copy number variation(CNV)in single cancer cells.