Evaluation of Two New Methods for DNA Extraction of “Legal High” Plant Species

Ryan, Angelique L.; O’Hern, P B S Cassandra; Elkins, Kelly M.

doi:10.1111/1556-4029.14478

Cited by 8 publications

(5 citation statements)

References 17 publications

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“…They also concluded that the QIA-GEN DNeasy kit is not a preferred method when dealing with large numbers of samples due to the long time that is required to complete the isolation. Our initial MicroGEM DNA concentration results were similar to previous research such as described by Ryan et al [39], which showed a very low recovered DNA concentration (less than 3 ng/µl) when using MIGROGEM on different plant tissues. After optimisation, we substantially increased the concentration to over 15 ng/µl and up to 78 ng/µl using MicroGEM.…”

Section: Discussionsupporting

confidence: 90%

Comparing DNA isolation methods for forest trees: quality, plastic footprint, and time-efficiency

Guillardín,

MacKay

2023

Plant Methods

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Background Genetic and genomic studies are seeing an increase in sample sizes together with a wider range of species investigated in response to environmental change concerns. In turn, these changes may come with challenges including the time and difficulty to isolate nucleic acids (DNA or RNA), the sequencing cost and environmental impacts of the growing amount of plastic waste generated in the process. Pseudotsuga menziesii var. menziesii (Mirbel) Franco (PM), Tsuga heterophylla (Raf.) Sarg. (TH) and Thuja plicata Donn ex D.Don (TP) are conifer species found in diverse woodlands both as natives and naturalized exotics. Our study was carried out whilst investigating their genetics to understand their population structure and potential for adaptation. Results In the present study, we compared two different DNA isolation methods, i.e., spin-column DNeasy plant mini kit (QIAGEN), and temperature-driven enzymatic cocktail Plant DNA Extraction (MicroGEM). The quantity of recovered DNA and the quality of DNA were assessed along with the plastic footprint and time needed for three tree species. Both methods were optimised and proven to provide enough DNA for each studied species. The yield of DNA for each method depended on the species: QIAGEN showed higher yield in P. menziesii and T. heterophylla, while T. plicata recovered similar amount of DNA for both methods. The DNA quality was investigated using DNA barcoding techniques by confirming species identity and species discrimination. No difference was detected in the PCR amplification of the two barcoding loci, (rbcL and trnH-psbA), and the recovered sequences between DNA isolation methods. Measurement of the plastic use and the processing time per sample indicated that MicroGEM had a 52.64% lower plastic footprint and was 51.8% faster than QIAGEN. Conclusions QIAGEN gave higher yields in two of the species although both methods showed similar quality results across all species. However, MicroGEM was clearly advantageous to decrease the plastic footprint and improve the time efficiency. Overall, MicroGEM recovers sufficient and reliable DNA to perform common downstream analyses such as PCR and sequencing. Our findings illustrate the benefits of research and efforts towards developing more sustainable methods and techniques to reduce the environmental footprint of molecular analyses.

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Section: Discussionsupporting

confidence: 90%

Comparing DNA isolation methods for forest trees: quality, plastic footprint, and time-efficiency

Guillardín,

MacKay

2023

Plant Methods

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“…Sinensis). At present, some studies have reported various of novel methods of medicinal plants identi cation, such as barcode-based melting curve analysis, DNA labeling or 1 HNMR metabolite ngerprinting and so on 26,27 . Although these methods can accurately identify species, they require specialized knowledge, statistical analysis, and complex instruments.…”

Section: Discussionmentioning

confidence: 99%

“…The extracted DNA was quanti ed using A260/280 ratio by QuickDrop™ Spectrophotometer, and all the values were greater than 1.80. To evaluate the sensitivity of the LAMP method, we diluted MO genomic DNA by a 10-fold dilution method, with a concentration range of 1 × 10 − 3 pg/µl to 1 × 10 3 pg/µl 26 . According to the commonly used genes encoding ITS, ITS2, psbA-trnH, matK, and rbcl published by GenBank, we selected the ITS2 gene sequence (No: AB715224.1) as the target gene for the primer sequence after referenced articles and used Blast analysis in NCBI (http://www.ncbi.nlm.nih.gov/BLAST).…”

Section: Herbal Sample Collection and Dna Extractionmentioning

confidence: 99%

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Rapid and reliable distinguish of Morinda officinalis How by loop-mediated isothermal amplification (LAMP) assay

Wang

Sui

Han

et al. 2020

Preprint

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The medicinal plant Morinda officinalis How (MO), especially the root, has been frequently used in traditional medicines around the world as an herbal drug for treating variable human disorders and diseases. Various adulterations of MO were found for economic or production limitations. However, authentication of MO from its adulterants by LAMP has not yet been established. The present study introduces a commercially available nucleic acid amplification method, loop-mediated isothermal amplification (LAMP) assay for the distinguish of MO from its adulterants. In this method, we combined DNA barcodes technology to design 2 pairs independent LAMP primers, which based on the internal transcribed spacer 2 (ITS2) sequence of MO’s nuclear ribosomal DNA. Our results showed that the LAMP could amplify the samples as expected and successfully identify target MO, and the limit for DNA template preciseness was verified as 1 × 10− 1 pg/µl. All the visual or real-time turbidity detection was performed within 60 min at approximately 63 °C. The result showed that the LAMP assay and primers we designed have high accuracy and efficiency for the differentiation of MO and its adulterants. Our results illustrated that the proposed low-cost, fast and reliable LAMP assay without the need for expensive equipment or specialized techniques could be a good way for MO rapid authentication.

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“…2019 GC-MS analysis of fifty samples of medicinal herbs collected from herb shops located in different parts of Iran [ 286 ]; overview of over 400 performance and image enhancing drugs confiscated in Italy in the period 2017–2019 [ 287 ]; evaluation of DNA extracted from the forensically relevant “legal high” plant species: Ipomoea purpurea, Artemisia absinthium, Mitragyna speciosa, Datura stramonium, and Papaver somniferum [ 288 ]; 2020 review of the types of drugs that may be illegally added in health food according to their pharmacological activities, advances in detection technologies for illegal drugs and future development prospects [ 289 ]; 2021 review of the significance of medicinal plants in forensic investigations to detect criminal offenses [ 290 ]; review of toxicological aspects and analytical methods for twelve plant specimens (Areca catechu, Argyreia nervosa, Ayahuasca, Catha edulis, Datura stramonium, Lophophora williamsii, Mandragora officinarum, Mitragyna speciosa, Piper methysticum Forst, Psilocybe, Salvia divinorum and Tabernanthe iboga) [ 291 ].…”

Section: Routine and Improved Analyses Of Abused Substancesmentioning

confidence: 99%