For each pooled extraction, the V4 region of the 16 S ribosomal RNA (16 S rRNA) gene was amplified in triplets using the universal primers 515 F and 806 R adapted with linker regions and barcoded sequences used for dual-indexing14. Platinum SuperFi DNA Polymerase (Thermo Fisher ...
Storage experiments were performed to mimic the storage and transfer of patient samples. Tissue samples were placed in PCR tubes and immersed in 200 µl of either RNA Later (Thermo Fisher, AM7020), DNA Shield (Zymo Research, R1100), TE buffer (pH 8, Promega, V6231), or PBS (pH 7.4...
(SG Medical Inc., Seoul, Korea). RNA was simultaneously extracted from 13 mL of serially diluted viral stocks of each strain using the three extraction systems. mRT-qPCR assay was performed using Allplex™ SARS-CoV-2 Assay kits. Ct values above 36 were not considered for evaluating ...
Up to now, the blood is still the best common source of biomarkers, which can provide impor- tant information for diagnosing disease, judging prognosis, and monitoring physiological metabolism [2,5–8]. Unfortunately, the blood collection procedure often involves or triggers tissue damage, ...
MicroRNA (miRNA) transfection is known to degrade target mRNAs and to decrease mRNA expression. In contrast to the notion that most of the gene expression alterations caused by miRNA transfection involve downregulation, they often involve both up- and downregulation; this phenomenon is thought to be...