To compare their ability to tune endogenous gene expression, we targeted the repressor systems to the Esrrb gene in a mESC line, where the gene is homozygously tagged with P2A-mCherry (Fig. 2a)36. We then used flow cytometry to quantify repression at the single-cell level. Previous work ...
(b) PCR analysis of cDNA from CHO and 2B1 cells or no cDNA (H2O as control) used primers as indicated in a. Products were analyzed by agarose gel electrophoresis and EtBr staining. (c) RNA was isolated from CHO (open bars) and 2B1 (filled bars) cells grown in media supplemented with...
We used a SYBR Green quantitative reverse transcriptase PCR (qRT-PCR) miRNA array to measure 84 miRNAs—chosen based on their presence in serum, plasma, and other extracellular bodily fluids, and their differential regulation in disease, including diabetes and heart disease (Additional File 2: ...
To quantify the ROS production in protoplasts, the H2DCFDA fluorescence signal after correction by Histogram Matching was used for plotting (Miura, 2020). For protoplast-based cell death assays, statistical analyses were performed with the Graphpad Prism 7.0 software (https://www.graphpad.com/). ...
Real-time PCR(often called qPCR) is usually conducted to quantify the absolute amount of a target sequence or to compare relative amounts of a target sequence between samples. This technique monitors the amplification of the target in real time via a target-specific fluorescent signal emitted duri...
3b). To confirm the interaction between miR-17-5p and the 3’UTR of KCNMA1, we used AGO2 immunoprecipitation [46, 47] to isolate the AGO2 protein and associated RNA following transfection with miR-17-5p mimic. This resulted in a clear increase in the KCNMA1-specific RT-PCR signal ...
3C). For MiD51, the sequence electropherogram of the PCR-amplified genomic region showed overlapping peaks (Fig. 3C), indicating the presence of heterozygous mutations in the different alleles. AltMiD51 was completely undetectable both by Western blotting (Fig. 3D) and absolute quantification (...
Microfluidic quantitative PCR for antibiotic resistance genes Remaining DNA from a subset of samples was used for microfluidic qPCR (MFQPCR) to quantify various antibiotic-resistance genes (ARGs) including those for beta-lactams, quinolones, and vancomycin resistance as previously described [26]. BioMark...
Apolipoprotein E4 (APOE4) is the strongest known genetic risk factor for late-onset Alzheimer’s disease (AD). Conditions of stress or injury induce APOE expression within neurons, but the role of neuronal APOE4 in AD pathogenesis is still unclear. Here
Real-time PCR was used to quantify SMN transcripts as described by Tiziano et al18 Primers SMN_mgb-F and SMN_mgb-R were used to amplify the SMN1 and SMN2 genes; the specific probes for SMN1 and SMN2 are listed in Table 1. GAPDH served as the internal control (primers GAPDH_abs-F...