GSM5799802: 6D2 IAA 10mg/L rep1; Mus musculus; RNA-Seq
On the morning of collection, estrous cyclicity was determined and ovaries were collected from mice in diestrus. Immediately after euthanasia, antral follicles were meticulously dissected from the ovaries based on size (220–400 µm) using watchmaker's forceps. Antral follicles (30-40 follicles per mouse) were dissected, snap frozen in liquid nitrogen, and then stored at –80 °C for RNA sequencing. Total RNA was isolated from the antral follicles using a RNeasy Micro kit (Qiagen, Inc., Valencia, California) following the manufacturer's instructions. The RNAseq libraries were prepared with Illumina's 'TruSeq Stranded mRNAseq Sample Prep kit' (Illumina). Read 1 aligns to the ANTISENSE strand. The libraries were pooled; quantitated by qPCR and sequenced on one SP lane for 101 cycles from one end of the fragments on a NovaSeq 6000. Fastq files were generated and demultiplexed with with the bcl2fastq v2.20 Conversion Software (Illumina). Adaptors have been trimmed from the 3'-end of the reads. Sequence of adaptors used to make the libraries (used the first 33nt for adaptor trimming). Adaptor sequence in read1: AGATCGGAAGAGCACACGTCTGAACTCCAGTCACNNNNNNNNATCTCGTATGCCGTCTTCTGCTTG (NNNNNNNN= 8 nt index).