AI Article Synopsis
- Extracting high-quality RNA from pancreatic tumors is difficult due to high ribonuclease concentrations, and existing methods often require time-consuming stabilization that can worsen RNA degradation.* -
- A newly optimized protocol offers a fast and inexpensive way to isolate high-quality RNA from mouse pancreatic tumors by using liquid nitrogen and guanidinium thiocyanate-chloroform extraction.* -
- The new method achieved a significant RNA Integrity Number (9.0), making it suitable for RNA sequencing (RNAseq) and quantitative PCR (qPCR) applications.*
Article Abstract
Extraction of high-quality RNA from pancreatic tumors for sequencing purposes is technically challenging, as the pancreas is an organ rich in ribonucleases. The majority of the established RNA isolation protocols for use with primary pancreatic tissue involve perfusion of RNA stabilizing reagent into the pancreatic tissue to protect RNA integrity before extraction. However, the additional time needed for this procedure can actually lead to further RNA degradation. We optimized a protocol suitable for high quality RNA isolation from mouse pancreatic tumors that is a simple, fast, and inexpensive modification of existing methods, combining the use of liquid nitrogen and guanidinium thiocyanate-chloroform extraction. Through this procedure, the mean RNA Integrity Number value obtained for RNA isolated from pancreatic tumors was 9.0, and was reproducibly suitable for RNAseq and qPCR.•a protocol suitable for high quality RNA isolation from mouse pancreatic tumors as well as normal pancreas•combining the use of liquid nitrogen and guanidinium thiocyanate-chloroform extraction.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7897710 | PMC |
| http://dx.doi.org/10.1016/j.mex.2020.101163 | DOI Listing |
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