Plant and metagenomic DNA extraction of mucilaginous seeds
The pulp surrounding the seeds of some fruits is rich in mucilage, carbohydrates, etc. Some seeds are rich in proteins and polyphenols. Fruit seeds, like cacao (Theobroma cacao) and cupuassu (Theobroma grandiflorum), are subjected to fermentation to develop flavor. During fermentation, ethanol is pr...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2014-01-01
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| Series: | MethodsX |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016114200628 |
| _version_ | 1811213255718207488 |
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| author | Simone N.M. Ramos Marcela M. Salazar Gonçalo A.G. Pereira Priscilla Efraim |
| author_facet | Simone N.M. Ramos Marcela M. Salazar Gonçalo A.G. Pereira Priscilla Efraim |
| author_sort | Simone N.M. Ramos |
| collection | DOAJ |
| description | The pulp surrounding the seeds of some fruits is rich in mucilage, carbohydrates, etc. Some seeds are rich in proteins and polyphenols. Fruit seeds, like cacao (Theobroma cacao) and cupuassu (Theobroma grandiflorum), are subjected to fermentation to develop flavor. During fermentation, ethanol is produced [2–6]. All of these compounds are considered as interfering substances that hinder the DNA extraction [4–8]. Protocols commonly used in the DNA extraction in samples of plant origin were used, but without success. Thus, a protocol for DNA samples under different conditions that can be used for similar samples was developed and applied with success. The protocol initially described for RNA samples by Zeng et al. [9] and with changes proposed by Provost et al. [5] was adapted for extracting DNA samples from those described. However, several modifications have been proposed:
•
Samples were initially washed with petroleum ether for fat phase removal.
•
RNAse was added to the extraction buffer, while spermidin was removed.
•
Additional steps of extraction with 5 M NaCl, saturated NaCl and CTAB (10%) were included and precipitation was carried out with isopropanol, followed by washing with ethanol. |
| first_indexed | 2024-04-12T05:43:55Z |
| format | Article |
| id | doaj.art-2668bf7c884746c38bef0f05fd3dcef8 |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-04-12T05:43:55Z |
| publishDate | 2014-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-2668bf7c884746c38bef0f05fd3dcef82022-12-22T03:45:31ZengElsevierMethodsX2215-01612014-01-011C22522810.1016/j.mex.2014.09.005Plant and metagenomic DNA extraction of mucilaginous seedsSimone N.M. Ramos0Marcela M. Salazar1Gonçalo A.G. Pereira2Priscilla Efraim3Department of Food Technology, School of Food Engineering, University of Campinas, BrazilLaboratory of Genomic and Expression, Institute of Biology, University of Campinas, BrazilLaboratory of Genomic and Expression, Institute of Biology, University of Campinas, BrazilDepartment of Food Technology, School of Food Engineering, University of Campinas, BrazilThe pulp surrounding the seeds of some fruits is rich in mucilage, carbohydrates, etc. Some seeds are rich in proteins and polyphenols. Fruit seeds, like cacao (Theobroma cacao) and cupuassu (Theobroma grandiflorum), are subjected to fermentation to develop flavor. During fermentation, ethanol is produced [2–6]. All of these compounds are considered as interfering substances that hinder the DNA extraction [4–8]. Protocols commonly used in the DNA extraction in samples of plant origin were used, but without success. Thus, a protocol for DNA samples under different conditions that can be used for similar samples was developed and applied with success. The protocol initially described for RNA samples by Zeng et al. [9] and with changes proposed by Provost et al. [5] was adapted for extracting DNA samples from those described. However, several modifications have been proposed: • Samples were initially washed with petroleum ether for fat phase removal. • RNAse was added to the extraction buffer, while spermidin was removed. • Additional steps of extraction with 5 M NaCl, saturated NaCl and CTAB (10%) were included and precipitation was carried out with isopropanol, followed by washing with ethanol.http://www.sciencedirect.com/science/article/pii/S2215016114200628PlantSeedsCupuassuDNA extractionPCRMetagenomic |
| spellingShingle | Simone N.M. Ramos Marcela M. Salazar Gonçalo A.G. Pereira Priscilla Efraim Plant and metagenomic DNA extraction of mucilaginous seeds MethodsX Plant Seeds Cupuassu DNA extraction PCR Metagenomic |
| title | Plant and metagenomic DNA extraction of mucilaginous seeds |
| title_full | Plant and metagenomic DNA extraction of mucilaginous seeds |
| title_fullStr | Plant and metagenomic DNA extraction of mucilaginous seeds |
| title_full_unstemmed | Plant and metagenomic DNA extraction of mucilaginous seeds |
| title_short | Plant and metagenomic DNA extraction of mucilaginous seeds |
| title_sort | plant and metagenomic dna extraction of mucilaginous seeds |
| topic | Plant Seeds Cupuassu DNA extraction PCR Metagenomic |
| url | http://www.sciencedirect.com/science/article/pii/S2215016114200628 |
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