GSM6594570: RNA expression, Tendinopathy; Homo sapiens; RNA-Seq
Tendinopathy tissue was placed in a 2-ml microcentrifuge tube, NP40 lysis buffer was added, and the mixture was incubated on ice for 5 min. Then, the mixture was filtered through a 70-µm filter and transferred to a new 2-ml microcentrifuge tube. After gradient centrifugation, the supernatant was removed. PBS, bovine serum albumin (BSA), and an RNase inhibitor were added and incubated on ice for 5 min. Repeated centrifugation resulted in a pure single-nuclei suspension. The obtained nuclear suspension was incubated in a transposase mixture that entered the nuclei and preferentially fragmented DNA in open chromatin regions. Oligonucleotides containing an Illumina P5 sequence, a 16-nt 10x barcode and a spacer sequence were then released. Barcoded, full-length preamplified cDNA was further amplified via PCR to generate sufficient mass for gene expression library construction.