Comparison of some DNA extraction methods from monovarietal must and wines

Abstract

The methods applied for DNA extraction from must and wine samples with monovarietal origin are presented and discussed aiming to prove the quality of extracted DNA and its good properties for further use in molecular tests. In the present research were compared four different DNA extraction methods from must and wine samples obtained from eleven V. vinifera varieties (five grapevine varieties for white wines and six grapevine varieties for red wines, respectively). Taking into consideration the amounts of obtained DNA, the concentrations and purities of the final DNA extracts, were stood out two modified methods. For all must samples, very efficient was the second method, which allowed obtaining a mean value of 87.9 ng µl-1 for the DNA concentration with 1.55 purity. Among the tested procedures, for monovarietal wine samples, the fourth method proved to be the most efficient which brought a mean value of 64.7 ng μl-1 for DNA concentration with 1.66 purity. This method adequate for wine samples involves two CTAB solution treatments and the RNase treatment applied before DNA resuspension. The DNA from must and wine extracts and the DNA from leaves of the corresponding grapevine varieties were amplified with five specific microsatellite primers (VVS2, VVMD27, VVMD32, VrZAG79 and VrZAG62). The aspects of pattern profiles were analysed in parallel and proved that the extracted DNA was suitable for amplification with these specific V. vinifera primers. The two selected extraction procedures are considered good for research purposes and ensure obtaining of good-quality extracted DNA from musts and one-year old wines.

Miniaturized Microscale Solid-Phase Extraction-Based Module for Highly Efficient DNA Extraction and Purification from Grapevine Samples

“…12 The main limitations of such analysis are related to the complexity of the vegetable sample, the loss of DNA integrity throughout food processing, 13 the risk of crosscontamination during various steps of DNA analysis, 14 and the presence of compounds that prevent DNA detection, such as PCR inhibitors. 15 Therefore, efficient DNA extraction and purification are crucial for the success of DNA-based methods. 15 Likewise, there is increasing interest in fast and reliable methods with high sensitivity and multiplexing possibilities.…”

“…15 Therefore, efficient DNA extraction and purification are crucial for the success of DNA-based methods. 15 Likewise, there is increasing interest in fast and reliable methods with high sensitivity and multiplexing possibilities. 16 In this sense, micro-total analysis systems (μTASs), also known as lab-on-a-chip, aim to completely integrate multiple laboratory processes in a single miniaturized platform.…”

“… 15 Therefore, efficient DNA extraction and purification are crucial for the success of DNA-based methods. 15 …”

Integrated Approach from Sample-to-Answer for Grapevine Varietal Identification on a Portable Graphene Sensor Chip

“…Pereira et al, 2011, reported a DNA wine extraction method starting with a 10 mL sample that resulted in a 3 ng DNA yield by 2-propanol precipitation, enzyme treatment, phenol and chloroform extraction, and several washes . Onache et al, 2021, showed the efficiency of DNA yield from different wine samples ranging from 18 to 85 ng/μL using Qiagen DNeasy Plant Kit . It should be taken into account that not all reportedly successful methods could be applied to any wine samples.…”

“… 25 Onache et al, 2021, showed the efficiency of DNA yield from different wine samples ranging from 18 to 85 ng/μL using Qiagen DNeasy Plant Kit. 26 It should be taken into account that not all reportedly successful methods could be applied to any wine samples. DNA amplification is frequently performed for the enrichment of the total DNA of a particular species by the use of universal primers for such species.…”