GSM3269917: IPN_miRNA_NAWD4; Mus musculus; miRNA-Seq
The IPN and MHb were dissected by a single midline circular tissue punch (0.75 mm diameter). Punches from 4 animals were pooled for each sample and total RNA was isolated using a miRVANA miRNA isolation kit (Ambion). Small RNAs (18-30 nt) were separated from the sample by denaturing gel electrophoresis. The small RNA pool was used to generate miRNA-Seq libraries and the remaining RNAs (> 30 nt) were used to generate stranded mRNA-Seq libraries. Small RNA libraries were synthesized using a modified protocol for Illumina TruSeq Small RNA Cloning (April 2014, www.zamorlab.umassmed.edu/protocols/). mRNA-Seq libraries were synthesized using the TruSeq Stranded mRNA Library Prep kit (Illumina) according to manufacturer's instructions. All libraries were gel-purified, assessed by Fragment Analyzer (Advanced Analytical, AATI) and their concentrations were quantified using KAPA Library Quantification Kits (KAPA Biosystems). miRNA-Seq libraries were pooled and single-end (75 cycles) sequencing was performed. mRNA-Seq libraries were pooled and paired-end (76 cycles x 2) sequencing was performed.
GEO Accession
GSM3269917