PMC:7464876 / 16077-16951 JSONTXT

{"target":"http://pubannotation.org/docs/sourcedb/PMC/sourceid/7464876","sourcedb":"PMC","sourceid":"7464876","source_url":"https://www.ncbi.nlm.nih.gov/pmc/7464876","text":"2.4. Quantitative Reverse Transcription PCR (qRT-PCR) Analysis\nTotal mRNAs were isolated from cells using the peqGold TriFast (Peqlab Biotechnologie, Darmstadt, Germany) and the Direct-zol™ RNA MiniPrep kit (Zymo Research, Irvine, CA, USA). The mRNA was digested with 5 U RNase-free, recombinant DNase I (Roche Diagnostics, Rotkreuz, Switzerland), and cDNA synthesis was performed by using the qScript cDNA Synthesis Kit (Quanta Biosciences, Beverly, MA, USA). qRT-PCR reactions were conducted as described [40]. For data analysis, the comparative cycle threshold method (CT) was used, and data were reported as the fold change normalized to an endogenous reference gene (β-actin). To quantify the ERGIC-53 transcript levels, the primers 5′- CAGATCAAATTCGAGTAGCACCA-3′ and 5′-AATATTCCAACACCATTCCACAGA-3′ were used. All other primer pairs have been described previously [29].","divisions":[{"label":"title","span":{"begin":0,"end":62}}],"tracks":[{"project":"2_test","denotations":[{"id":"32806600-32017353-20296203","span":{"begin":870,"end":872},"obj":"32017353"}],"attributes":[{"subj":"32806600-32017353-20296203","pred":"source","obj":"2_test"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"2_test","color":"#a0ec93","default":true}]}]}}