Quantifiler duo dna quantification kit

The DNA methylation of the cells from the cell cultures was analyzed using MagMeDIP qPCR Kit (Diagenode, Denville, NJ, USA) according to the manufacturer’s protocol. After precipitation and DNA extraction, DNA concentration was measured, using Pico488 dsDNA quantification reagent (Lumiprobe, Cockeysville, MD, USA). The standard curve was prepared from human DNA quantified by Quantifiler™ Duo DNA Quantification Kit (Thermo Fisher, Waltham, MA, USA). The global DNA methylation was quantified as the percentage of DNA immunoprecipitated using C-me antibodies (Diagenode, Denville, NJ, USA) to the input amount of DNA, based on the concentration results. The site-specific DNA methylation was analyzed in Real-Time PCR, using Fast SYBR Green Master Mix (Thermo Fisher, Waltham, MA, USA). The primer sequences are presented in Table 1. The promoter region of the IL10 gene was the target of primers. The calculation of % of the input was done according to the manufacturer’s protocol. The following formula was used:
where, CtInput—Ct value of 10% Input; CtCme—Ct value of precipitated DNA using Cme antibody.

DNA methylation was analyzed using the MagMeDIP qPCR Kit (Diagenode, Denville, NJ, USA) according to protocol. After precipitation and DNA extraction, the DNA concentration was measured using Pico488 dsDNA quantification reagent (Lumiprobe, Hannover, Germany). The standard curve was prepared using human DNA quantified by the Quantifiler™ Duo DNA Quantification Kit (ThermoFisher Scientific). Global DNA methylation was calculated as the percentage of DNA immunoprecipitated using C^me antibodies (Diagenode) to the input amount of DNA. The site-specific DNA methylation was analyzed in real-time PCR using the Fast SYBR Green Master Mix (ThermoFisher Scientific). The primer sequences are presented in Table 1. The calculation of percent of input was performed according to the manufacturer protocol:
CtIN—Ct value of 10% input; CtIP—Ct value of immunoprecipitated samples].

According to manufacturer recommended protocols, reference profiles and standard bulk mixture samples were extracted using the QIAamp DNA Ivestigator Kit (QIAGEN, Germantown, MD, USA) and quantified using Quantifiler^® Duo DNA Quantification kit (ThermoFisher Scientific, Carlsbad, CA, USA) on the Applied Biosystems’ 7500 real-time PCR instrument (ThermoFisher Scientific, Carlsbad, CA, USA). One nanogram of DNA extracts were amplified using the GlobalFiler^TM amplification kit (ThermoFisher Scientific, Carlsbad, CA, USA) at 29 PCR cycles. Probabilistic genotyping of donor references profiles and complex equimolar 2-6 person mixtures was conducted using STRmix™ v2.8 and EuroForMix v3.1.0 (Quantitative LR MLE based) according to the known NOC.