GRO-Seq analysis of in vitro activated B cells in early G1

Purpose: To detect the on-going transcription in early G1 B cells Methods: Primary B cells were isolated and in vitro activated by LPS, IL4 and Rp105, followed by nocodazole arrest and release. The early G1 population was then sorted and the nuclei was isolated for GRO-Seq. The sequenced reads were processed using Burrows–Wheeler Aligner (BWA) followed by HOMER. Results: The nascent transcription and the convergent transcription were determined in both unsynchronized and sorted early G1 population Conclusions: Convergent transcription is detected at Igh switch regions in early G1 cells. Two samples for unsynchronized B cells, and two samples for sorted early G1 cells.