Genome copies per µl RNA template were calculated based on a quantified standard RNA, where absolute quantification was done by the QX200 Droplet Digital PCR System in combination with the 1-Step RT-ddPCR Ad
For real-time qPCR, total RNA was isolated from cultured cells using the PureLink RNA Mini Kit (Thermo Fisher). Complementary DNA (cDNA) was generated using the SuperScript VILO cDNA Synthesis Kit (Thermo Fisher). ddPCR was carried out on the reverse transcribed cDNA using QX200™ ddPCR™ ...
hTERT splicing was determined with RT- ddPCR assays. h Telomerase enzyme activity (ddTRAP with 50 cell equivalents added to the assay) in Calu6 cells with and without NOVA1 (n = 3). Student's t test set at *p < 0.05 for significance). Data are expressed as means and standard error ...
The relative expression of the mutant and wild-type alleles was determined using 2-step RT–PCR. Primers were designed to target exons 4 and 5 of the MLLT1 gene (NM_ 005934). Complementary DNA was synthesized using the SuperScript VILO cDNA kit from Invitrogen. The manufacture protocol was ...
RT-qPCR run. First, the absolute concentration of the 07V063 stock was determined by droplet digital PCR (ddPCR) using the QX600 Droplet Digital PCR System (Bio-Rad Laboratories, Hercules, CA, USA) with the One-Step RT-ddPCR Advanced Kit for Probes (Bio-Rad Laboratories, Hercules, CA, ...