GSM3244797: d0RNAseq_rep1; Mus musculus; RNA-Seq
Total RNA was isolated from 2-5 million trypsinized cells using trizol extraction. polyA+ mRNA was isolated using the NEBNext NEBNext® Poly(A) mRNA Magnetic Isolation Module using 5 ug of total RNA as input. Isolated mRNA was reverse-transcribed using Superscript III and actinomycin D to inhibit template switching. Second-strand synthesis was performed using the NEBNext Ultra Directional RNA Second Strand Synthesis Module. Library preparation and NEBNext® ChIP-Seq Library Prep Master Mix Set for Illumina. A USER enzyme treatment was performed following adaptor ligation to specifically degrade the second strand and allow a stranded analysis. Libraries were amplified for 10-15 cycles of PCR using Q5 polymerase and NEBNext multiplex oligos.
GEO Accession
GSM3244797