Proven performance—over 10 years of clinical real-time PCR instrument manufacturing Designed with the clinic in mind—interactive diagnostic instrument with short run times, minimal maintenance, and use of existing plastic consumables Enhanced security—auditing an...
And 50–80-pM probes were added in the hybridization buffer to incubate for one night. For RNA loading, 5S rRNA was used as control. qRT-PCR analysis of miRNA was carried out as described [64], using miRNA 1st Strand cDNA Synthesis Kit (Vazyme Biotech, China) and miRNA Universal SYBR ...
polyA-tailed RNA isolation: two rounds of poly(A) enrichment were conducted using DynaBeads mRNA Direct Purification Kit (Thermo Fisher Scientific) following the manufacturer’s protocol with some modifications. In brief, 200 ml of Dynabeads were washed with 200 ml of lysis/binding buffer and ...
Quantitative RT-PCR Total RNA was isolated from cells using the Rneasy Mini Kit (74106, Qiagen). For cDNA synthesis, total RNA was transcribed using PrimeScript (DRR047A, Takara, Dalian, China). The levels of specific RNAs were measured using the ABI 7900 real-time PCR machine and the Fa...
5′-aacgcttcacgaatttgcgt-3′ (antisense); All operations were carried out at least 3 times. 2.8. Western blotting Treated cells were extracted with total protein with RIPA lysis buffer (Beyotime) to measure the protein expressions. Western blotting was performed according to standard procedures,...
Quantitative real-time PCR (qRT–PCR) was used to detect levels of HCG18, miR-29a/b, and mRNAs. MTT, colony formation, wound healing and Transwell assays were used to evaluate cell proliferation, migration and invasion, respectively. A luciferase reporter assay was utilized to evaluate NF-κB...
Dimethyl sulfoxide (1-10%) has been shown to accelerate strand renaturation and is believed to give the nucleic acid thermal stability against depurination. As a PCR cosolvent, DMSO may help improve yields, especially in long PCR. Uses
Organoids were harvested in RLT buffer at 0, 1, or 6 hours post the addition of actinomycin D and processed for RT-PCR. Acknowledgments We would like to acknowledge the Flow Cytometry Core at Stony Brook University. We would like to acknowledge the Research Histology Core Laboratory at Stony...
real-time PCR was carried out using the LightCycler System (Roche Applied Science, Mannheim, Germany). PCR was set up at 2.5 mM MgCl2, 10 μM of each primer (Eurofins Genomics, USA), 4 μl of recover DNA and 5 µl of Master Mix (KAPA Biosystems) in a final volume of ...
FCRL5 gene expression was analyzed via qRT-PCR (BlazeTaq™ SYBR® Green qPCR Mix2.0), using GAPDH as a reference. This investigation adhered to the Declaration of Helsinki and received approval from the Ethics Committee of West China Hospital at Sichuan University (reference: 2018–061). ...