Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 213.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 247.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 247.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 247.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 247.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Deprecated: Implicit conversion from float 247.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534
Warning: imagejpeg(C:\Inetpub\vhosts\kidney.de\httpdocs\phplern\27510798
.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 Mol+Biol+Rep
2016 ; 43
(10
): 1165-78
Nephropedia Template TP
gab.com Text
Twit Text FOAVip
Twit Text #
English Wikipedia
Identification of extracellular miRNA in archived serum samples by
next-generation sequencing from RNA extracted using multiple methods
#MMPMID27510798
Gautam A
; Kumar R
; Dimitrov G
; Hoke A
; Hammamieh R
; Jett M
Mol Biol Rep
2016[Oct]; 43
(10
): 1165-78
PMID27510798
show ga
miRNAs act as important regulators of gene expression by promoting mRNA
degradation or by attenuating protein translation. Since miRNAs are stably
expressed in bodily fluids, there is growing interest in profiling these miRNAs,
as it is minimally invasive and cost-effective as a diagnostic matrix. A
technical hurdle in studying miRNA dynamics is the ability to reliably extract
miRNA as small sample volumes and low RNA abundance create challenges for
extraction and downstream applications. The purpose of this study was to develop
a pipeline for the recovery of miRNA using small volumes of archived serum
samples. The RNA was extracted employing several widely utilized RNA isolation
kits/methods with and without addition of a carrier. The small RNA library
preparation was carried out using Illumina TruSeq small RNA kit and sequencing
was carried out using Illumina platform. A fraction of five microliters of total
RNA was used for library preparation as quantification is below the detection
limit. We were able to profile miRNA levels in serum from all the methods tested.
We found out that addition of nucleic acid based carrier molecules had higher
numbers of processed reads but it did not enhance the mapping of any miRBase
annotated sequences. However, some of the extraction procedures offer certain
advantages: RNA extracted by TRIzol seemed to align to the miRBase best;
extractions using TRIzol with carrier yielded higher miRNA-to-small RNA ratios.
Nuclease free glycogen can be carrier of choice for miRNA sequencing. Our
findings illustrate that miRNA extraction and quantification is influenced by the
choice of methodologies. Addition of nucleic acid- based carrier molecules during
extraction procedure is not a good choice when assaying miRNA using sequencing.
The careful selection of an extraction method permits the archived serum samples
to become valuable resources for high-throughput applications.