Τίτλος:
The Effect of Genomic DNA Contamination on the Detection of Circulating
Long Non-Coding RNAs: The Paradigm of MALAT1
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
The presence of contaminating gDNA in RNA preparations is a frequent
cause of false positives in RT-PCR-based analysis. However, in some
cases, this cannot be avoided, especially when there are no exons-intron
junctions in the lncRNA sequences. Due to the lack of exons in few of
long noncoding RNAs (lncRNAs) and the lack of DNAse treatment step in
most studies reported so far, serious questions are raised about the
specificity of lncRNA detection and the potential of reporting
false-positive results. We hypothesized that minute amounts of gDNA
usually co-extracted with RNA could give false-positive signals since
primers would specifically bind to gDNA due to the lack of junction. In
the current study, we evaluated the effect of gDNA and other forms of
DNA like extrachromosomal circular DNAs (eccDNAs) contamination and the
importance of including a DNAse treatment step on lncRNAsexpression.As a
model, we have chosen as one of the most widely studied lncRNAs in
cancer namely MALAT1, which lacks exons. When we tested this hypothesis
in plasma and primary tissue samples from NSCLC patients, our findings
clearly indicated that results on MALAT1 expression are highly affected
by the presence of DNA contamination and that the DNAse treatment step
is absolutely necessary to avoid false positive results.
Συγγραφείς:
Markou, Athina N.
Smilkou, Stavroula
Tsaroucha, Emilia and
Lianidou, Evi
Περιοδικό:
DIAGNOSTIC ONCOLOGY
Λέξεις-κλειδιά:
MALAT1; non-long coding RNA; DNAse treatment; cfRNA; tumor biomarkers;
false positive results; gDNA
DOI:
10.3390/diagnostics11071160
