title: GSM5577734: Drug_D7_LD9R_Luminal; Rattus norvegicus; RNA-Seq
library_construction_protocol: single cell: mammary gland single-cell suspensions were stained with Live/Dead aqua stain the following markers: CD24, CD29, CD31, and CD45 in PBE (PBS with 0.5% BSA and 2 mM EDTA, for 30 min at 4oC). Basal, luminal cells, and leukocytes were sorted into lysis buffer and RNA was extracted using teh AllPrep DNA/RNA mini kit (Qiagen)  Total RNA was extracted using the RNeasy Mini Kit (Qiagen). bulk: The total RNA was measured using the Agilent 2100 Bioanalyzer. RNA-seq libraries were prepared using Clontech Low Input mRNA Library (Clontech SMARTer) v4 kit from less than 10ng of purified total RNA according to the manufacturer's protocol. The concentrations of finished dsDNA library were measured using the Qubit Fluorometer, the size of library fragment was measured by Agilent TapeStation 2200, and RT-qPCR for adapted library molar concentration measurement according to manufacturer's protocols. Uniquely indexed libraries were pooled in equimolar ratios and sequenced on an Illumina NextSeq500 with Single-End 75bp (SE75) reads by the Dana-Farber Cancer Institute Molecular Biology Core Facilities. RNA libraries were prepared for sequencing using standard Illumina protocols