RNA Extraction and RT-PCR Analysis

Total RNA was extracted using Trizol® Reagent (Invitrogen). RT-PCR was performed as described previously [7 (link)] using a reaction kit containing M-MLV reverse transcriptase (Roche) and ProTaq® DNA polymerase (Roche).The primers used for c-Maf, CCND2, ITGB7 and GAPDH were described previously [7 (link)]. The thermal cycle conditions used were as follows: 1 cycle at 94 °C for 5 min; 35 cycles at 94 °C for 30 s; 55 °C for 30 s; 72 °C for 30 s; and 1 cycle at 72 °C for 10 min.

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Sun T., Xu Y., Xu Z., Cao B., Zhang Z., Wang Q., Kong Y, & Mao X. (2021). Inhibition of the Otub1/c-Maf axis by the herbal acevaltrate induces myeloma cell apoptosis. Cell Communication and Signaling : CCS, 19, 24.

Publication 2021

Trizol® Reagent M-MLV reverse transcriptase ProTaq® DNA polymerase

Ccnd2 Dna polymerase Gapdh Itgb7 Primers Reverse transcriptase Rt pcr Trizol

Corresponding Organization :

Other organizations : Guangzhou University of Chinese Medicine, Soochow University, First Affiliated Hospital of Soochow University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression levels of c-Maf, CCND2, ITGB7 and GAPDH

control variables

Thermal cycle conditions: 1 cycle at 94 °C for 5 min; 35 cycles at 94 °C for 30 s; 55 °C for 30 s; 72 °C for 30 s; and 1 cycle at 72 °C for 10 min.
Reaction kit components: M-MLV reverse transcriptase (Roche) and ProTaq® DNA polymerase (Roche)
Primers used for c-Maf, CCND2, ITGB7 and GAPDH as described previously [7 (link)]

controls

No positive or negative controls were explicitly mentioned in the provided information.

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