Quantifying AMBRA1 Gene Expression

The total RNAs were extracted from cells using total RNA kit (Tiangen, Beijing, China). cDNA was synthesized using ReverTra Ace RT Master Mix (Toyobo, Osaka, Japan). qPCR assay was performed to assess the relative abundances of AMBRA1 and GAPDH mRNAs using specific primers (Table 3), stained by SYBR Green (Tiangen) on the CFX96 real-time PCR system (Bio-Rad, California, USA). The relative abundances of AMBRA1 were normalized to that of GAPDH, using the 2^ΔΔCt method [15 (link)]. All data were obtained from three independent experiments.

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Zhao B., Yang Y., Cun B, & Chen P. (2021). AMBRA1 attenuates the proliferation of uveal melanoma cells. Open Medicine, 17(1), 1-14.

Publication 2021

total RNA kit ReverTra Ace RT Master Mix SYBR Green CFX96 real-time PCR system

Ambra1 Assay Cdna Gapdh Mrnas Primers Sybr green

Corresponding Organization : Kunming Medical University

Other organizations : Center for Excellence in Molecular Cell Science, Chinese Academy of Sciences

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Variable analysis

independent variables

Not explicitly mentioned

dependent variables

Relative abundances of AMBRA1 mRNA
Relative abundances of GAPDH mRNA

control variables

Not explicitly mentioned

controls

Positive control: Not mentioned
Negative control: Not mentioned

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