GSM4227746: A12 NET-seq Rep 2; Saccharomyces cerevisiae; OTHER
Cells were lysed via 10 1 minute grinds at maximum amplitude in a mikro dismembrator II. The grinding cup was chilled in liquid nitrogen for 1 minute between each grind. NET-seq libraries were prepared as in Clarke, et. al., NETSeq reveals heterogeneous nucleotide incorporation by RNA polymerase I, PNAS, 2018. 115 (50), E11633-E11641. In brief,  Pol I elongation complexes were isolated from the cell lysate via immunoprecipitation with anti-HA magnetic beads. Associated RNA transcripts were then isolated using an acidic phenol-chloroform extraction. The RNA transcripts were then 3' appended with a DNA linker and reverse transcribed to produce complementary DNAs (cDNAs). The cDNAs were circularized and amplified by PCR to produce high-throughput sequencing libraries. NET-seq
GEO Accession
GSM4227746