Faculty
Schulich School of Medicine and Dentistry
Supervisor Name
Matthew Grol
Keywords
tendon, RNA isolation, histology, sectioning, mouse
Description
Tendons join muscle to bone and are essential for posture and movement. Tenocytes are the resident tendon cells. Isolating RNA from small animal models such as mice can be difficult, especially from tendon, as much of the time, they are extremely small samples. Downstream analysis such as qPCR and especially RNA sequencing require greater quantity and quality of RNA than is often achievable. RNA degradation is extremely rapid following tendon isolation, with RNA quality decreasing as time increases between the collection of tendons and RNA isolation. Even tendons whose RNA was isolated immediately following tendon isolation had imperfect quality. The goals for this project were to first, find and optimize a reproducible method for isolating RNA from mouse tendons for downstream qPCR and next generation sequencing that is replicable for people with varying degrees of experience, and second, optimize a method of sectioning mouse joints for the examination of tendon tissue structure.
Acknowledgements
Thank you to all members of the Grol Lab, and in particular, Mayeesha Khan. Thank you to Melika Meshkat, Mayeesha Khan, Ali Naeij, Dawn Bryce, and Dr. Matt Grol for their help, guidance, and support with this project
Thank you to CMHR and the USRI program for this opportunity and funding, and thank you to the Schulich School of Medicine and Dentistry.
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial 4.0 License
Document Type
Poster
Optimizing RNA Isolation and Histology Protocols for Characterization of Preclinical Models of Tendon Disease
Tendons join muscle to bone and are essential for posture and movement. Tenocytes are the resident tendon cells. Isolating RNA from small animal models such as mice can be difficult, especially from tendon, as much of the time, they are extremely small samples. Downstream analysis such as qPCR and especially RNA sequencing require greater quantity and quality of RNA than is often achievable. RNA degradation is extremely rapid following tendon isolation, with RNA quality decreasing as time increases between the collection of tendons and RNA isolation. Even tendons whose RNA was isolated immediately following tendon isolation had imperfect quality. The goals for this project were to first, find and optimize a reproducible method for isolating RNA from mouse tendons for downstream qPCR and next generation sequencing that is replicable for people with varying degrees of experience, and second, optimize a method of sectioning mouse joints for the examination of tendon tissue structure.