A Simple and Rapid System for DNA and RNA Isolation from Diverse Plants Using Handmade Kit

Abstract

This protocol describes a rapid DNA and RNA extraction method for plant tissues. Hexadecyltrimethylammonium bromide (CTAB), sodium chloride (NaCl), tris base, and ethylenediaminetetraacetic acid (EDTA) are the main components of the extraction buffer. In contrast to all previously reported protocols, this extraction method does not require any stock solutions. This isolation buffer is potential of extracting both DNA and RNA simultaneously. Depending on the purpose of the project, the corresponding steps can be slightly altered to obtain either DNA or RNA. The big advantage of this method is to use general laboratory chemicals to make a powerful extraction buffer, resulting in high quality and quantity nucleic acid. Also, CTAB in this buffer is capable of isolating nucleic acid from recalcitrant plants enriched in secondary metabolites. Importantly, this method is recommended for the projects at which isolating nucleic acid in a short time is of crucial importance. This method probably is usable for all plant tissues and takes about an hour.

Patterns of gene expression in pollen of cotton (Gossypium hirsutum) indicate downregulation as a feature of thermotolerance

“…Total RNA was isolated from approximately 15–50 mg of tissues, using a CTAB method optimized by Masoomi‐Aladizgeh et al. (2016), followed by applying a RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). Briefly, 500 μl of the extraction buffer in addition to 25 μl of beta‐mercaptoethanol were added to the ground pollen, and the samples were incubated at 65°C for 5 min.…”

Genome survey sequencing of wild cotton (Gossypium robinsonii) reveals insights into proteomic responses of pollen to extreme heat

“…Total genomic DNA was extracted from approximately 100 mg young leaf tissue using a combination of a method described in Masoomi‐Aladizgeh et al ( 2016 ) and a DNeasy Plant Mini Kit (Qiagen). Briefly, a lysis buffer for DNA extraction was prepared as described in Masoomi‐Aladizgeh et al ( 2016 ).…”

“…Total genomic DNA was extracted from approximately 100 mg young leaf tissue using a combination of a method described in Masoomi‐Aladizgeh et al ( 2016 ) and a DNeasy Plant Mini Kit (Qiagen). Briefly, a lysis buffer for DNA extraction was prepared as described in Masoomi‐Aladizgeh et al ( 2016 ). The lysis buffer (1200 µl) in addition to β‐mercaptoethanol (50 µl) was added to the ground leaf and samples were incubated at 65°C for 10 min.…”