Genes, Vol. 12, Pages 1617: Isolation of RNA from Acute Ischemic Stroke Clots Retrieved by Mechanical Thrombectomy

Genes, Vol. 12, Pages 1617: Isolation of RNA from Acute Ischemic Stroke Clots Retrieved by Mechanical Thrombectomy Genes doi: 10.3390/genes12101617 Authors: Vincent M. Tutino Sarah Fricano Kirsten Frauens Tatsat R. Patel Andre Monteiro Hamid H. Rai Muhammad Waqas Lee Chaves Kerry E. Poppenberg Adnan H. Siddiqui Mechanical thrombectomy (MT) for large vessel acute ischemic stroke (AIS) has enabled biologic analyses of resected clots. While clot histology has been well-studied, little is known about gene expression within the tissue, which could shed light on stroke pathophysiology. In this methodological study, we develop a pipeline for obtaining useful RNA from AIS clots. A total of 73 clot samples retrieved by MT were collected and stored in RNALater and in 10% phosphate-buffered formalin. RNA was extracted from all samples using a modified Chemagen magnetic bead extraction protocol on the PerkinElmer Chemagic 360. RNA was interrogated by UV–Vis absorption and electrophoretic quality control analysis. All samples with sufficient volume underwent traditional qPCR analysis and samples with sufficient RNA quality were subjected to next-generation RNA sequencing on the Illumina NovaSeq platform. Whole blood RNA samples from three patients were used as controls, and H&E-stained histological sections of the clots were used to assess clot cellular makeup. Isolated mRNA was eluted into a volume of 140 µL and had a concentration ranging from 0....
Source: Genes - Category: Genetics & Stem Cells Authors: Tags: Article Source Type: research