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2014 ; 11
(7
): 817-28
Nephropedia Template TP
gab.com Text
Twit Text FOAVip
Twit Text #
English Wikipedia
Capture and Amplification by Tailing and Switching (CATS) An ultrasensitive
ligation-independent method for generation of DNA libraries for deep sequencing
from picogram amounts of DNA and RNA
#MMPMID24922482
Turchinovich A
; Surowy H
; Serva A
; Zapatka M
; Lichter P
; Burwinkel B
RNA Biol
2014[]; 11
(7
): 817-28
PMID24922482
show ga
Massive parallel sequencing (MPS) technologies have paved the way into new areas
of research including individualized medicine. However, sequencing of trace
amounts of DNA or RNA still remains a major challenge, especially for degraded
nucleic acids like circulating DNA. This together with high cost and time
requirements impedes many important applications of MPS in medicine and
fundamental science. We have established a fast, cheap and highly efficient
protocol called 'Capture and Amplification by Tailing and Switching' (CATS) to
directly generate ready-to-sequence libraries for MPS from nanogram and picogram
quantities of both DNA and RNA. Furthermore, those DNA libraries are
strand-specific, can be prepared within 2-3 h and do not require preliminary
sample amplification steps. To exemplify the capacity of the technique, we have
generated and sequenced DNA libraries from hundred-picogram amounts of
circulating nucleic acids isolated from human blood plasma, one nanogram of
mRNA-enriched total RNA from cultured cells and few nanograms of
bisulfite-converted DNA. The approach for DNA library preparation from minimal
and fragmented input described here will find broad application in diverse
research areas such as translational medicine including therapy monitoring,
prediction, prognosis and early detection of various human disorders and will
permit high-throughput DNA sequencing from previously inaccessible material such
as minute forensic and archeological samples.