Chunk #13 — Materials and Methods — RNA Isolation and cDNA Synthesis

Source

PASSPORT-seq: A Novel High-Throughput Bioassay to Functionally Test Polymorphisms in Micro-RNA Target Sites.

Embedded

yes

Text

Transfected cells were incubated for 48 h, lysed in situ and total RNA isolated using a RNeasy® purification kit with the optional DNase treatment (Qiagen, Germantown, MD, United States). RNA was quantified using the Quant-iTTM RNA Broad Range kit (Thermo Fisher Scientific, Waltham, MA, United States) and cDNA synthesized from 800 ng of total RNA using the QuantiTect® Reverse Transcription kit (Qiagen, Germantown, MD, United States).