Hot firepol solisgreen qpcr mix reagent

Animals were synchronized by bleaching and plated as L1 larvae on NGM agar plates seeded with E. coli HT115 (EV), which were kept at 20 °C. Animals were collected at the L4 stage and frozen in liquid nitrogen. TRIzol Reagent (ThermoFisher Scientific, #15596018) was used to extract RNA. cDNA synthesis was done with the QuantiTect Reverse Transcription Kit (Qiagen, #205313) and qRT-PCR reactions were run with HOT FIREPol SolisGreen qPCR Mix-reagent (Solis BioDyne, #08-46-00001) using the CFX384 machine (Bio-Rad). qRT-PCR data were normalized to the expression of cdc-42 and pmp-3. qRT-PCR oligos used in this study are provided in Supplementary Information file 1, Table S2. qRT-PCR experiments were performed with three biological replicates, with three technical replicates for each biological replicate. Analysis of ribosomal subunit expression was performed once with each folr-1 mutant, and the analysis of folr-1 RNAi efficiency in N2 and folr-1 OE strains was performed twice with similar results. Statistical significances were analyzed by using Student’s t-test and one-way ANOVA.

Animals were synchronized by bleaching and plated as L1 larvae on NGM agar plates seeded with E. coli HT115 (EV). Plates were kept at 20 °C. Animals were collected at the L4 stage and frozen in liquid nitrogen. TRIzol Reagent (Thermo Fisher Scientific, #15596018) was used to extract RNA. cDNA (complementary DNA) synthesis was done with the QuantiTect Reverse Transcription Kit (Qiagen, #205313) and quantitative reverse transcription polymerase chain reaction (qRT-PCR) reactions were run with the HOT FIREPol SolisGreen qPCR Mix-reagent (Solis BioDyne, #08-46-00001) using the CFX384 machine (Bio-Rad). qRT-PCR data were normalized to the expression of cdc-42 and pmp-3. qRT-PCR oligos used in this study are provided in Supplementary Material File 1, Table S3. qRT-PCR experiments were performed with three biological replicates, with three technical replicates for each biological replicate. Statistical significances were analyzed using an unpaired t-test.