ABA-responsive Gene Expression Analysis

Two-week-old seedlings were incubated in liquid MS medium with or without 50 μM ABA for 3 h. Total RNAs was extracted using the RNAiso Plus reagent from Takara (Otsu, Japan). The cDNAs were synthesized from 1 μg of total RNAs using the PrimeScript RT-PCR reagent Kit of Takara. qRT-PCR was performed using the Bio-Rad CFX96 real-time PCR detection system and SYBR Premix Ex Taq II from Takara. ACTIN2/8 was used as an internal control. RT-PCR was conducted with the gene-specific primers described by Zhang et al. (2018) (link).

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Li X., Kong X., Huang Q., Zhang Q., Ge H., Zhang L., Li G., Peng L., Liu Z., Wang J., Li X, & Yang Y. (2018). CARK1 phosphorylates subfamily III members of ABA receptors. Journal of Experimental Botany, 70(2), 519-528.

Publication 2018

RNAiso Plus reagent PrimeScript RT-PCR reagent Kit CFX96 real-time PCR detection system SYBR Premix Ex Taq II

Cdnas Gene Primers Rnas Rt pcr Seedlings

Corresponding Organization : Sichuan University

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Variable analysis

independent variables

ABA treatment (presence or absence of 50 μM ABA)

dependent variables

Gene expression levels measured by qRT-PCR

control variables

ACTIN2/8 as an internal control gene

controls

Positive control: Seedlings incubated in liquid MS medium without ABA
Negative control: Not explicitly mentioned

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