Rapid and simple method for DNA extraction from plant and algal species suitable for PCR amplification using a chelating resin Chelex 100

Abstract

A DNA extraction method using Chelex 100 is widely used for bacteria, Chlamydomonas, and animal cell lines, but only rarely for plant materials due to the need for additional time-consuming and tedious steps. We have modified the Chelex 100 protocol and successfully developed a rapid and simple method of DNA extraction for efficient PCR-based detection of transgenes from a variety of transgenic plant and algal species. Our protocol consists of homogenizing plant tissue with a pestle, boiling the homogenized tissue in a microfuge tube with 5% Chelex 100 for 5 min, and centrifuging the boiled mixture. The supernatant, which is used for PCR analysis, was able to successfully amplify transgenes in transgenic tobacco, tomato, potato, Arabidopsis, rice, strawberry, Spirodela polyrhiza, Chlamydomonas, and Porphyra tenera. The entire DNA extraction procedure requires \15 min and is therefore comparable to that used for bacteria, Chlamydomonas, and animal cell lines.

Botanical and Genetic Identification Followed by Investigation of Chemical Composition and Biological Activities on the Scabiosa atropurpurea L. Stem from Tunisian Flora

“…Moreover, a genetic characterization using bio-molecular tools were applied to plant fresh material in order to confirm genus and species identification. Briefly, DNA from Scabiosa stems, freshly collected was extracted using the chelating resin Chelex 100 [ 52 ]. The extraction eluate was subjected to a polymerase chain reaction (PCR) amplification of a Chloroplastic DNA fragment, using the species- and group-specific primers “ycf6F” and “psbMR”, targeting for the petN-psbM intergenic spacer region [ 53 ].…”

Accumulated lipids rather than the rigid cell walls impede the extraction of genetic materials for effective colony PCRs in Chlorella vulgaris

“…In addition, Modifications of the DNA extraction using Chelex-100 [ 14 ] involve additional mechanical homogenization for higher plants [ 29 ] or high temperature incubation in ethanol following homogenization for Chlamydomonas reinhardtii and Arabidopsis thaliana [ 30 ]. The protocol presented here dispenses with these procedures, involves fewer steps and equipment.…”

Extraction of DNA from lichen-forming and lichenicolous fungi: a low-cost fast protocol using Chelex

“…Although this technique has been known for almost 30 years (Singer-Sam et al 1989), is commonly used in human forensics (Walsh et al 1991) and has also been successfully used in other eukaryotes (e.g. Pedersen et al 2006;HwangBo et al 2010), this study appears to be the first time it has been reported for use in lichen-forming and lichenicolous fungi.…”

“…It has been successfully applied in a wide range of eukaryotic organisms (e.g. Pedersen et al 2006; Strange et al 2009; HwangBo et al 2010; Casquet et al 2012) including some non-lichenized fungi (e.g. Zhang et al 2010; Turan et al 2015).…”