Targeted sequencing of reporter transcripts with variable GA-multivalency and intron content
Targeted sequencing of reporter transcripts with variable GA-multivalency and intron content
Samples were collected by trypsinising, spinning down, washing in PBS, spinning down and snap freezing on dry ice. Cells were grown in DMEM+Glutamax+10% FBS and seeded such that they achieved 70% confluency on the morning of the experiment, with each sample being derived from 2 wells of a 6 well plate. Cells were either transfected, dox-induced, and sometimes drug treated for the period of time described in the accession description and sample names. RNA extractions were performed from cell pellets using the Maxwell RSC simplyRNA kit (Promega) using a Maxwell RSC Instrument (Promega). Superscript IV RT using a barcoded RT primer specific to the reporter transcript. Specific amplification of the barcoded sequences using nested PCR. Final library amplification using indexed i5/i7 Illumina primers.