Paramagnetic cellulose DNA isolation improves DNA yield and quality among diverse plant taxa

Abstract

• Premise of the study: The chemical diversity of land plants ensures that no single DNA isolation method results in high yield and purity with little effort for all species. Here we evaluate a new technique originally developed for forensic science, based on MagnaCel paramagnetic cellulose particles (PMC), to determine its efficacy in extracting DNA from 25 plant species representing 21 families and 15 orders.• Methods and Results: Yield and purity of DNA isolated by PMC, DNeasy Plant Mini Kit (silica column), and cetyltrimethylammonium bromide (CTAB) methods were compared among four individuals for each of 25 plant species. PMC gave a twofold advantage in average yield, and the relative advantage of the PMC method was greatest for samples with the lowest DNA yields. PMC also produced more consistent sample purity based on absorbance ratios at 260:280 and 260:230 nm.• Conclusions: PMC technology is a promising alternative for plant DNA isolation.

Nucleic acid purification from plants, animals and microbes in under 30 seconds

“…The use of cellulose for DNA purification is commercially available and is claimed to have improved performance over silica-based DNA purification methods [ 58 , 59 ]. DNA purification using cellulose has been previously reported to be achieved by coaggregating or adsorbing the DNA to the cellulose in the presence of various chemicals, including chaotropic salts [ 60 ], ethanol [ 23 , 61 ], and high salt concentrations and/or crowding agents such as polyethylene glycol [ 11 , 62 , 63 ], which destabilise the DNA structure and facilitate its interaction with the cellulose fibres.…”

Automated DNA extraction using cellulose magnetic beads can improve EGFR point mutation detection with liquid biopsy by efficiently recovering short and long DNA fragments

“…A newly automated system using cellulose magnetic beads for capturing DNA was recently developed and is now commercially available. Because a matrix secondary fibril-associated cellulose was applied to DNA purification in this system, a wide range of DNA sizes (from 100 bp to 50 Kb) could be isolated [ 26 ], leading to high DNA yield. In addition to isolating a large quantity of DNA, this system enabled us to improve the efficiency of EGFR mutation detection.…”

Rapid (30-second), equipment-free purification of nucleic acids using easy-to-make dipsticks

“…Although a number of substrates can capture nucleic acids [7][8][9][10] , the novelty of our discovery is the transformation of a low-cost cellulose filter paper into a convenient dipstick that can rapidly purify nucleic acids from crude samples and elute them directly into the amplification mix without any pipetting. The dipstick method uses a mechanism of purification that is different from those of other systems; it is thought to take advantage of the differences between nucleic acids and other compounds in regard to their capture and retention kinetics by the cellulose matrix 5 .…”

“…Commercially available nucleic acid purification methodologies typically involve capturing nucleic acids in the presence of high concentrations of chaotropic salts (e.g., paramagnetic beads or spin columns) and, in comparison to dipstick purifications, are more expensive and more time consuming. They require several pipetting steps and, in the case of spin columns, require the use of centrifuges 5 , 7 , 8 . Adaptions of these technologies have been developed using a variety of nucleic acid binding substrates, including diatomaceous earth 9 , aluminum oxide 10 and fusion-5 filters 19 , but again, these adaptions are more complicated to set up and require several pipetting steps, making them less practical for high-throughput or remote field applications.…”