A 3-µl volume of cDNA (1:20 dilution) was used in a 10 µl quantitative-PCR reaction using SsoAdvanced Universal SYBR Green Supermix (BioRad, cat no. 1725271) and 250 nM primers (Supplementary Table 1). Parallel tbb-2 quantitative PCR reactions were run for each sample for ...
The slides were immersed in 70% ethanol for 5 min, dried at 37 °C for 15 min and then stained in SYBR Gold nucleic acid gel stain (Invitrogen, S11494) for 30 min in the dark. Analysis of neutral comet assay plus radiation was performed 24 h after 5 Gy ionizing ...
Oligonucleotide products were then resolved on a 2% agarose gel and visualized after staining with Sybr Gold (Invitrogen, Carlsbad, CA). The wild-type allele resulted in amplification of a 1300-bp product and the null allele a 600-bp product [55]. Histological analysis of circumvallate ...
Quantitative real-time RT-PCR analysis was performed using SYBR® Fast qPCR Mix (TaKaRa, Shiga, Japan) and a Agilent Technologies Stratagene Mx3000p Real-Time System (Agilent, USA) with SYBR Green to determine the mRNA expression level of a gene of interest. A reference gene, glyceraldehyde...
Power SYBR Green PCR Thermo Fisher Cat# 4367659 Phusion high-fidelity DNA Polymerase New England Biolabs Cat# M0530S HiScribe T7 Quick High Yield RNA Synthesis kit New England Biolabs Cat# E2050S Monarch RNA Cleanup kit New England Biolabs Cat# T2030S RNA 6000 Nano Kit Agilent Cat# 5067-15...
Table 2 on the LightCycler 480 thermocycler (Roche Diagnostics). For calculating target gene levels relative to the house keeping geneATPsyn-Cf6the ΔΔct method including efficiency correction was used. Determination of ctvalues was done by calculating the second derivative maximum of the SYBR green...
RT-qPCR assays consisting of three biological and three technical replicates were performed on a CFX96 (Bio–Rad) using the SYBRGreen PCR Supermix kit (Bio–Rad Laboratories). Banana MaRPS476 and tomato SlActin77 genes were adopted as the reference genes. Transcript levels were calculated ...
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qPCR was conducted using Universal SYBR qPCR Master Mix (Biosharp, BL697A). NbActin was used for internal control. Primers used are listed in Additional file 1: Table S3. DNA extraction and qPCR Total DNA was extracted from plant leaves by the CTAB method. For detection of AtGeBP T-DNA...
RT-PCR analysis was performed with a CFX real-time PCR instrument (Bio-Rad) and SYBR Green mixture (Roche). The relative expression of the genes was quantified with the 2−ΔΔCT calculation, using UBIQUITIN as the reference housekeeping gene for the expression analyses. The enrichment of ...