Zr duet dna rna miniprep plus

Extraction of total nucleic acids after enzymatic removal of the proteins was followed by fractionation of DNA and RNA with the standard TRIzolTM protocol (Rio et al., 2010). Both fraction of nucleic acid aqueous phase (RNA) and chloroform-water interphase separately ethanol precipitated, again, once more fractionated through binding onto Zymo-SpinTM columns (ZYMO-RESEARCH CORP, Irvine, CA, USA) and elution columns or by chloroform extraction (www.zymoresearch.com accessed on 2 March 2021, ZR-Duet ™ DNA/RNA MiniPrep Plus catalog number D7003). The free RNA was separated, and the fraction that remains bound to the DNA further purified after DNase digestion and purification with Zymo-SpinTM columns (ZYMO-RESEARCH CORP, Irvine, CA, USA).

DNA and RNA were isolated from the same piece of tissue or cell pellet using ZR-Duet DNA/RNA Miniprep Plus (Zymo Research, D7001). If only DNA was extracted, the DNeasy Blood & Tissue Kit (Qiagen, 69581) was used, and if only RNA was extracted, the RNeasy Mini Kit protocol (Qiagen, 74104) was used. DNA and RNA quality were measured using a Nanodrop spectrophotometer (Thermo-Scientific). DNA concentration was determined using a Qubit fluorometer (Thermo Fisher Scientific, Q32850 and Q32851). RNA integrity was analyzed and quantified using a 4200-Tapestation (Agilent).