Introducing nano-scale quantitative polymerase chain reaction

• Published in: Biochemical and biophysical research communications Vol. 506; no. 4; pp. 923 - 926
• Main Authors: Dyrstad, Sissel E., Tusubira, Deusdedit, Knappskog, Stian, Tronstad, Karl J., Røsland, Gro V.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 02.12.2018
• Subjects: 60 APPLIED LIFE SCIENCES; GENES; Nano-scale qPCR; POLYMERASE CHAIN REACTION; POLYMERASES; qPCR; TaqMan technology; qPCR; Nano-scale qPCR; TaqMan technology
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2018.10.111

We show that the quantitative PCR reaction volume can be reduced significantly compared to the standard procedures, without compromising data quality. By analyzing dilution series (100–10−7) we found that measurement in 1 μl reaction volume indeed gave valid results comparable to 10 μL, when using a routine pipetting robot. This may enable a significant cost reduction through cutback of both biological material as well as chemical reagents. •qPCR remain the gold-standard-method for measuring and validating gene expression levels and alterations.•qPCR reaction volume can be significantly reduced.•1 μL versus 10 μL total reaction volumes gave comparable results.•The reduction in reaction volume can greatly reduce sample use and reagent cost.