Illumina MiSeq paired end sequencing; Identification of a novel ESR1 transcript variant in breast cancer cells by RNA capture-sequencing
Identification of a novel ESR1 transcript variant in breast cancer cells by RNA capture-sequencing
Cultured breast cancer cells were harvested, and pellets were washed 3X with cold PBS solution before RNA extraction. Trizol / Chloroform extraction followed by isopropanol precipitation Total RNA samples (100 ng) were input in the RNA library construction using the KAPA RNA Hyper library prep kit. Protocol was modified based on the quality and quality of the samples. Customized adapters with unique molecular indexes (UMI) and Sample-specific dual-indexes primers were added to each library. The quantity of libraries was measured with Qubit and quality measured by TapStation Genomic DNA Assay.