Quantitative analysis of long non-coding RNAs

Briefly, cDNA was synthesized from total RNA using a PrimeScript RT reagent kit with gDNA Eraser (Takara, Shiga, Japan). Primers for lncRNAs were designed and synthesized. Then, qPCR assays were performed using a Bio-Rad iQ5 Multicolor Real-Time qRT-PCR Detection system (Bio-Rad, Hercules, CA, USA). The expression levels of lncRNAs were detected as previously described (21 (link)). Human β-actin was used as a housekeeping gene for normalization. The expression levels of lncRNAs were measured in terms of the cycle threshold (CT) and were then normalized to β-actin expression using the 2^−ΔΔCt method (21 (link)).

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Ren S., Li G., Liu C., Cai T., Su Z., Wei M., She L., Tian Y., Qiu Y., Zhang X., Liu Y, & Wang Y. (2016). Next generation deep sequencing identified a novel lncRNA n375709 associated with paclitaxel resistance in nasopharyngeal carcinoma. Oncology Reports, 36(4), 1861-1867.

Publication 2016

PrimeScript RT reagent kit with gDNA Eraser iQ5 Multicolor Real-Time qRT-PCR Detection system

Actin Assays Cdna Housekeeping gene Human Lncrnas Primers

Corresponding Organization :

Other organizations : Xiangya Hospital Central South University, Central South University

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Variable analysis

independent variables

Primers for lncRNAs

dependent variables

Expression levels of lncRNAs

control variables

Human β-actin (used as a housekeeping gene for normalization)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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