Improved microRNA quantification in total RNA from clinical samples.
microRNAs are small regulatory RNAs that are currently emerging as new biomarkers for cancer and other diseases. In order for biomarkers to be useful in clinical settings, they should be accurately and reliably detected in clinical samples such as formalin fixed paraffin embedded (FFPE) sections and blood serum or plasma. These types of samples represent a challenge in terms of microRNA quantification. A newly developed method for microRNA qPCR using Locked Nucleic Acid (LNA)-enhanced primers enables accurate and reproducible quantification of microRNAs in scarce clinical samples. Here we show that LNA-based microRNA qPCR enables biomarker screening using very low amounts of total RNA from FFPE samples and the results are compared to microarray analysis data. We also present evidence that the addition of a small carrier RNA prior to total RNA extraction, improves microRNA quantification in blood plasma and laser capture microdissected (LCM) sections of FFPE samples.
Andreasen
Ditte
D
Exiqon A/S, Skelstedet 16, DK-2950 Vedbaek, Denmark. ina@exiqon.com
Fog
Jacob U
JU
Biggs
William
W
Salomon
Jesper
J
Dahslveen
Ina K
IK
Baker
Adam
A
Mouritzen
Peter
P
D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D007834:Lasers; D035683:MicroRNAs / Q000032:analysis* / Q000097:blood / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction / Q000379:methods*
D005404:Fixatives; D035683:MicroRNAs; D005557:Formaldehyde