Turbo dna free dnase treatment and removal reagents

qRT-PCR was carried out on genomic DNA and RNA samples to determine relative copy number and relative RNA expression levels of transgenes, respectively. Genomic DNA was extracted from approximately 5.0 × 10⁶ cells using GeneJET Genomic DNA Purification Kit (Thermo Fisher Scientific). RNA was extracted from a minimum of 1.0 × 10⁶ cells using TRIzol™ Reagent (Thermo Fisher Scientific), followed by DNase treatment to remove contaminating DNA (TURBO DNA-free™ DNase Treatment and Removal Reagents, Thermo Fisher Scientific). cDNAs were synthesized from 1 μg of total RNAs using Maxima First Strand cDNA Synthesis Kit for RT-qPCR (Thermo Fisher Scientific). The qRT-PCR was performed in an Mx3005P qPCR System (Agilent Technologies) using Brilliant III Ultra-Fast SYBR^® Green QPCR Master Mix (Agilent Technologies) as described previously^{23 (link)}. Target regions were amplified using primers listed in Supporting Table S3 that were validated by melting curve analysis and agarose gel electrophoresis. Fold changes were calculated using the ∆∆CT method. Vinculin was used as normalizer for genomic DNA quantification^{23 (link)} and β-actin (ACTB) was used for RNA quantification since the expression of β-actin RNA/protein was considered constant and was not affected by temperature in CHO cells^{50 (link)}.