RNA-seq of negative control and KRT32-overexpressing Ker-CT cells
Release date:
2024-03-18
Description:
The RNA-seq experiment involved three replicate samples of negative control and KRT32-overexpressing Ker-CT cells. The cells were initially cultured for 24 hours in a 12-well plate with a cell count of 10^6. Total RNA was extracted using the RNAiso Plus kit (TAKARA, Japa). Eukaryotic mRNA enrichment was achieved using Oligo(dT) beads, and the enriched mRNA fragments were further fragmented using fragmentation buffer. These fragments were then reverse transcribed into cDNA using the NEBNext Ultra RNA Library Prep Kit for Illumina from New England Biolabs. After end-repair and addition of A bases to the double-stranded cDNA fragments, Illumina sequencing adapters were ligated. The ligated fragments underwent size selection via agarose gel electrophoresis and were then amplified through PCR. Finally, the resulting cDNA library was sequenced on an Illumina Novaseq6000 platform.
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Individuals & samples
Files
Submitter: Liu Hong / hongyue2519@hotmail.com
Organization: Shandong Provincial Hospital for Skin Diseases & Shandong Provincial Institute of Dermatology and Veneorology, Shandong First Medical University & Shandong Academy of Medical Sciences
Submission date: 2024-03-18
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HRA006924.xlsx