GSM2176504: SIRT7 RIP RNA, nucleus rep1; Homo sapiens; RIP-Seq
For the extraction of total SIRT7-RIP RNA, we first fractionated cell lysates into cytosolic and nuclear fractions and then performed RIP (RNA immunoprecipitation) experiment. Briefly, the lysates were incubated with anti-Flag M2 affinity gel to immunoprecipitate SIRT7 ribonucleoprotein complex (RNP) . Anti-Flag beads were washed extensively and then subjected to proteinase K digestion to release RNA from SIRT7 RNP. RNA associated with SIRT7 was then extracted by standard phenol-chloroform extraction followed with ethanol precipitation. Directional RNA-seq libraries were prepared from 25-100ng input RNA using the NEBNext Ultra Directional RNA Library Prep Kit for Illumina (New England Biolabs).
GEO Accession
GSM2176504