GSM4718652: F508delCFBE41o, siCTRL, vehicle (Urban_3_1_S9); Homo sapiens; RNA-Seq
Total RNA was isolated from cell lysate using the Quick-RNA Miniprep Kit (Zymo Research, R1054) according to the manufacturer's instructions, with additional on-column DNase treatment with the RNase-free DNase Set (Zymo) to remove contaminating genomic DNA 1 μg of total RNA was used with the Illumina TruSeq Stranded Total RNA sample preparation kit (Illumina, San Diego, CA, USA), following manufacturer's instruction. 3′ and 5' adapters were ligated to total input RNA. Reverse transcription followed by polymerase chain reaction (PCR) was used to create cDNA constructs. This process selectively enriches those fragments that have adapter molecules on both ends. PCR was performed with primers that anneal to the ends of the adapters. cDNA libraries were then purified using QIAquick PCR purification kit (Qiagen). The cDNA libraries were validated using KAPA Biosystems primer premix kit with Illumina-compatible DNA primers and Qubit 2.0 fluorometer. Quality was examined using Agilent Bioanalyzer Tapestation 2200. The cDNA libraries were pooled at a final concentration of 1.8pM. Cluster generation and 75 bp paired-end read sequencing was performed on Illumina NextSeq 500's (Illumina).
GEO
GSM4718652