Aug 24, 2022
Version 2
Wet Chemistry RNA Isolation Method for Mature Cassava Leaf Tissue V.2
This protocol is a draft, published without a DOI.
- 1Realizing Increased Photosynthetic Efficiency (RIPE)
Protocol Citation: Lynn Doran 2022. Wet Chemistry RNA Isolation Method for Mature Cassava Leaf Tissue. protocols.io https://protocols.io/view/wet-chemistry-rna-isolation-method-for-mature-cass-cfrqtm5wVersion created by Lynn Doran
Manuscript citation:
Behnam, Babak, Adriana Bohorquez-Chaux, Oscar Fernando Castaneda-Mendez, Hiroyuki Tsuji, Manabu Ishitani, and Luis Augusto Becerra Lopez-Lavalle. “An Optimized Isolation Protocol Yields High-Quality RNA from Cassava Tissues (Manihot Esculenta Crantz).” FEBS Open Bio 9, no. 4 (2019): 814–25. https://doi.org/10.1002/2211-5463.12561.
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 24, 2022
Last Modified: July 11, 2023
Protocol Integer ID: 69136
Keywords: RNA, RNA Extraction, Cassava
Abstract
RNA extraction protocol for cassava for RNA of sufficient quality and quantity to perform RT-PCR gene expression studies.
*Low recovery was achieved using Qiagen RNeasy kit. Typical recoveries using the mini protocol outlined here yielded 500-1300 ng/ul RNA from ~ 75 mg leaf tissue.*
Attachments
Safety warnings
- This protocol uses chemical fume hoods and centrifuges. Understand how to safely and appropriately use them before performing the protocol.
- Perform all steps within a fume hood and collect tips and tubes in the hazardous material collection bins. β-mercaptoethanol (β-ME) included in the extraction buffer is toxic, harmful to the environment and corrosive. Collect hazardous waste and submit to UIUC DRS following proper protocol.
- Some of the chemicals used in this protocol pose serious health risks. Please read all manufacturer safety data sheets before handling. UIUC personnel performing this protocol should be current on "Laboratory Safety", "Chemical Safety- An Introduction", and "Chemical Spills" Division of Research Safety training modules before performing this protocol.