Version 1
: Received: 8 March 2024 / Approved: 11 March 2024 / Online: 11 March 2024 (05:39:28 CET)
How to cite:
Ahmad, M. F.; Elias, M. H.; Mat Jin, N.; Abu, M. A.; Syafruddin, S. E.; Zainuddin, A. A.; Suzuki, N.; Abdul Karim, A. K. RNA Extraction from Cumulus Cells for Gene Analysis: The Current Strategy and Review of Literature. Preprints2024, 2024030573. https://doi.org/10.20944/preprints202403.0573.v1
Ahmad, M. F.; Elias, M. H.; Mat Jin, N.; Abu, M. A.; Syafruddin, S. E.; Zainuddin, A. A.; Suzuki, N.; Abdul Karim, A. K. RNA Extraction from Cumulus Cells for Gene Analysis: The Current Strategy and Review of Literature. Preprints 2024, 2024030573. https://doi.org/10.20944/preprints202403.0573.v1
Ahmad, M. F.; Elias, M. H.; Mat Jin, N.; Abu, M. A.; Syafruddin, S. E.; Zainuddin, A. A.; Suzuki, N.; Abdul Karim, A. K. RNA Extraction from Cumulus Cells for Gene Analysis: The Current Strategy and Review of Literature. Preprints2024, 2024030573. https://doi.org/10.20944/preprints202403.0573.v1
APA Style
Ahmad, M. F., Elias, M. H., Mat Jin, N., Abu, M. A., Syafruddin, S. E., Zainuddin, A. A., Suzuki, N., & Abdul Karim, A. K. (2024). RNA Extraction from Cumulus Cells for Gene Analysis: The Current Strategy and Review of Literature. Preprints. https://doi.org/10.20944/preprints202403.0573.v1
Chicago/Turabian Style
Ahmad, M. F., Nao Suzuki and Abdul Kadir Abdul Karim. 2024 "RNA Extraction from Cumulus Cells for Gene Analysis: The Current Strategy and Review of Literature" Preprints. https://doi.org/10.20944/preprints202403.0573.v1
Abstract
RNA extraction from human cumulus cells (CCs) is considered challenging due to low cell’s pop-ulation and small cell. The successful gene expression analysis from cumulus cells ultimately de-pends on an excellent concentration of Ribonucleic acid (RNA) yield by an appropriate technique. Our study aims to consolidate appropriate cumulus cell collection method, preparation and ex-traction techniques to ensure a good yield of RNA for better gene expression analysis. From the results obtained, the optimal strategy found involved mechanical denudation without an enzy-matic process, followed by the use of RNA stabilizer prior to RNA extraction. Subsequently, uti-lizing a smaller homogenizer, preferably manual pressure control with filter (Bio-Masher III®) is paramount for a pure and homogenize sample. Subsequently, during RNA extraction, the use of RNeasy micro kit and RNA carriers is recommended. These sample collection, preparation, and extraction techniques yielded an excellent-quality RNA concentration and successful gene expres-sion analysis. We also conducted a systematic review to consolidate our findings with current evidence of RNA extraction technique. Our results added a value as a current strategy for opti-mizing RNA extraction in CCs for experimental studies. Thus, our findings can contribute to for-mulating a better strategy for optimizing the RNA extraction in CCs for transcriptomic studies.
Medicine and Pharmacology, Obstetrics and Gynaecology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.