RNA Extraction and qRT-PCR Analysis

Fat body tissues were lysed using TRIzol reagent (Invitrogen), and total RNA was extracted according to a protocol described previously [50 (link)]. Then, single-stranded cDNA was synthesized using PrimeScript RT reagent Kit with gDNA Erase (TaKaRa) starting from 1 μg total RNA. The qRT-PCR of target genes was performed using SuperReal PreMix (Tiangen) in a StepOnePlus Fast Real-Time PCR system (Thermo Fisher Scientific), and each reaction was run in triplicate. Template concentration was normalized to an endogenous control ribosomal protein S7 (RPS7). The primers used for qRT-PCR are listed in the S1 Table.

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Wang M., Wang Y., Chang M., Wang X., Shi Z., Raikhel A.S, & Zou Z. (2022). Ecdysone signaling mediates the trade-off between immunity and reproduction via suppression of amyloids in the mosquito Aedes aegypti. PLoS Pathogens, 18(9), e1010837.

Publication 2022

TRIzol reagent gDNA Erase SuperReal PreMix

Cdna Fat body tissues Genes Primers Ribosomal protein s7 Trizol

Corresponding Organization : University of California, Riverside

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Expression levels of target genes

control variables

Endogenous control ribosomal protein S7 (RPS7) used to normalize template concentration

controls

Positive control: Not specified
Negative control: Not specified

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