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2017 ; 7
(1
): 11030
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Complete sequence-based pathway analysis by differential on-chip DNA and RNA
extraction from a single cell
#MMPMID28887473
van Strijp D
; Vulders RCM
; Larsen NA
; Schira J
; Baerlocher L
; van Driel MA
; Pødenphant M
; Hansen TS
; Kristensen A
; Mir KU
; Olesen T
; Verhaegh WFJ
; Marie R
; van der Zaag PJ
Sci Rep
2017[Sep]; 7
(1
): 11030
PMID28887473
show ga
We demonstrate on-chip, differential DNA and RNA extraction from a single cell
using a microfluidic chip and a two-stage lysis protocol. This method enables
direct use of the whole extract, without additional washing steps, reducing
sample loss. Using this method, the tumor driving pathway in individual cells
from a colorectal cancer cell line was determined by applying a Bayesian
computational pathway model to sequences obtained from the RNA fraction of a
single cell and, the mutations driving the pathway were determined by analyzing
sequences obtained from the DNA fraction of the same single cell. This combined
functional and mutational pathway assessment of a single cell could be of
significant value for dissecting cellular heterogeneity in tumors and analyzing
single circulating tumor cells.