real time fluorescent quantitative pcr instrument  (Bio-Rad)

Journal: Frontiers in Immunology

Article Title: Study on the spatiotemporal regulation of interferon-stimulated genes during Zika virus infection

doi: 10.3389/fimmu.2025.1702266

Figure Lengend Snippet: Effects of seven interferon-stimulated gene (ISG) knockdown on Zika virus (ZIKV) replication in A549 cells. A549 cells were transfected with small interfering RNA (siRNA) targeting candidate ISGs or non-targeting control siRNA for 48 h, followed by infection with ZIKV at a multiplicity of infection (MOI) of 1. The viral copy number in the supernatant was measured by quantitative real-time PCR (qRT-PCR) at 24 h post-infection. Data represent the mean ± SD from three or more independent experiments. Significance was determined using two-sided Student’s t -test. * p < 0.05; ** p < 0.005.

Article Snippet: A total of 1,000 ng of qualified total RNA was used to synthesize complementary DNA (cDNA) using the PrimeScriptTM RT kit (catalog no. RR047A; TaKaRa, Shiga, Japan) under the following reaction conditions: 37°C for 15 min and 85°C for 5 s. Using the synthesized cDNA as a template, detection was performed on a real-time fluorescent quantitative PCR instrument (LightCycler 96; Roche, Basel, Switzerland) with SYBR Green fluorescent dye (catalog no. RR420A; TaKaRa, Shiga, Japan).

Techniques: Knockdown, Virus, Transfection, Small Interfering RNA, Control, Infection, Real-time Polymerase Chain Reaction, Quantitative RT-PCR