Quantification of Oncogenic lncRNA Targets

We performed real-time quantitative polymerase chain reaction (RT-QPCR) using the ABI Prism 7700 Sequence Detector System (Applied Biosystems, Foster City, CA, USA) to determine the lncRNA level. Based on previous studies [11 (link),12 (link),13 (link),14 (link),15 (link),16 (link),17 (link),19 (link),20 (link),21 (link),22 (link),23 (link),24 (link),25 (link),30 (link)], we quantified the lncRNA targets maternally expressed gene 3 (MEG3), antisense non-coding RNA in the INK4 locus (ANRIL), long non-coding mega-cluster (lnc-MGC), metastasis-associated lung carcinoma transcript 1 (MALAT1), cancer susceptibility candidate 2 (CASC2), and taurine-upregulated gene 1 (TUG1). All primer and probe sequences were custom-designed (Applied Biosystems). Small RNA U6 (Applied Biosystems) was used as a house-keeping gene to normalize the lncRNA level [31 (link)]. Results were analyzed with Sequence Detection Software version 2.0 (Applied Biosystems) using the ΔΔCT method for relative quantitation, and results were expressed as copy number per 1000 copies of the housekeeping gene.

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Szeto C.C., So H., Poon P.Y., Luk C.C., Ng J.K., Fung W.W., Chan G.C., Chow K.M., Lai F.M, & Tam L.S. (2023). Urinary Long Non-Coding RNA Levels as Biomarkers of Lupus Nephritis. International Journal of Molecular Sciences, 24(14), 11813.

Publication 2023

Sequence Detection Software version 2.0

Cancer Gene Housekeeping gene Ink4 Lncrna Lung carcinoma Metastasis Non coding rna Primer Prism Real time polymerase chain reaction Susceptibility Taurine

Corresponding Organization : Chinese University of Hong Kong

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

LncRNA level

control variables

Small RNA U6 (Applied Biosystems) was used as a house-keeping gene to normalize the lncRNA level

controls

Positive control: None mentioned
Negative control: None mentioned

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