A Modified SDS – Based Method Applied for Extraction of High-Quality DNA from Raw Corn and Roasted Soybean

AbstractThe probability of contamination of non-transgenic varieties with genetically modified (GM) products increase as a result of global expansion of areas sown with transgenic crops. DNA-based methods as accurate, efficient and reliable methods are preferable for detection of GM material in raw or highly processed foods. Isolation of high quality DNA with a suitable and efficient DNA extraction protocol is crucial for getting precise results in DNA amplification. In this study, we performed modifications of previously known Sodium dodecyl sulfate (SDS)-based DNA extraction method regarding the incubation period, DNA pellet washing and addition of organic solvent extraction, to improve DNA quality and to reduce costs. Raw corn kernels and roasted soybean seed were used as samples. DNA was extracted following three protocols, modifications of Edwards protocol. The type of detergent used in raw corn sample did not cause significant effects on extracted DNA yield and purity, while in roasted soybean samples the 2% (w/v) SDS lysis buffer gave the highest DNA yield. The additional incubation step raised the DNA yield from raw corn for 121%, while the purest DNA from soybean sample was obtained using organic solvent extraction. Electrophoretic determination of DNA integrity showed varying degree of DNA smearing from roasted soybean. Contrary, all extraction protocols used on raw corn kernels produced a high molecular weight DNA. Thus, our in-house DNA extraction protocol is as efficient but more cost effective compared to commercial kits and can be used for raw corn, while the protocol for roasted soybean needs further improvement.

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DNA Extraction from Bronchial Aspirates for Molecular Cytology: Which Method to Take?

Analytical Cellular Pathology ◽

10.1155/2003/354796 ◽

2003 ◽

Vol 25 (2) ◽

pp. 83-88 ◽

Cited By ~ 7

Author(s):

Hans Jürgen Grote ◽

Viola Schmiemann ◽

Mario Sarbia ◽

Alfred Böcking

Keyword(s):

Success Rate ◽

Dna Extraction ◽

Extraction Method ◽

Globin Gene ◽

Extraction Methods ◽

High Quality ◽

High Molecular Weight Dna ◽

Dna Yield ◽

Extraction Protocol ◽

Dna Extraction Method

Objective: To date, there are only few systematic reports on the quality of DNA extracted from routine diagnostic cytologic specimens. It was the aim of the present study to evaluate the ability of 50% ethanol/2% carbowax (Saccomanno fixative) to preserve bronchial secretions with high quality genomic DNA as well as to compare different DNA extraction methods.Methods: DNA was extracted from 45 bronchial aspirates by four different extraction protocols. Beside DNA yield, DNA quality with regard to purity, integrity, and PCR success rate were investigated.Results: No fragmentation of sample DNA due to the fixative was detected. It was preserved as high molecular weight DNA. DNA yield, purity, and integrity were dependent on the DNA extraction method to some extend. Irrespective of the DNA extraction method the PCR success rate for amplification of β‐globin gene fragments (268, 536, and 989 bp) was 100%. Conclusions: A fixative containing 50% ethanol/2% carbowax preserves high quality DNA which is well suited for PCR‐based assays regardless of the extraction protocol used. The selection of the DNA extraction protocol has to be adjusted to the circumstances of application.

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Rnase A Enzyme Modification of Optimized SDS Protocol for DNA Extraction Suitable for Real-Time PCR Screening of GMOs

Macedonian Veterinary Review ◽

10.2478/macvetrev-2021-0028 ◽

2021 ◽

Vol 0 (0) ◽

Author(s):

Arita Sabriu-Haxhijaha ◽

Velimir Stojkovski ◽

Gordana Ilievska ◽

Dean Jankuloski ◽

Katerina Blagoevska

Keyword(s):

Real Time ◽

Dna Extraction ◽

Real Time Pcr ◽

Genetically Modified Crops ◽

Dna Amplification ◽

Rnase A ◽

High Quality ◽

Pcr Screening ◽

Dna Yield ◽

Corn Kernels

Abstract As the number of genetically modified crops increases rapidly, their accurate detection is significant for labelling and safety assessment. Currently, real-time PCR is the “golden standard” method for GMO detection. Hence, extraction of high quality DNA represents a crucial step for accurate and efficient DNA amplification. For GMO presence evaluation in the extracted DNA from raw corn kernels and roasted soybean, we used real-time PCR method, in consistent with the ISO17025 accreditation standards. As for DNA extraction, modified basic SDS protocol by adding RNase A enzyme in different steps of the protocol, with different time and temperature of incubation was used. The results showed as most suitable, the protocol where 10 µl of RNase A enzyme was added together with the lysis buffer at 65 °C for 30 minutes. Data for DNA yield and purity for roasted soybean was 469.6±3.3 µg/ml with A260/280 absorbance ratio 1.78±0.01. Suitability of DNA extracts for GMO analysis was assessed by screening for the presence of 35S promotor and Tnos terminator. Diluted extracts in concentrations 10, 1, 0.1, 0.01 and 0.0027 ng/µl, were tested in six replicates. Positive signal of amplification (LOD) was detected in all concentrations for both genetic elements in both matrices. The LOQ for 35S and Tnos for both matrices was 0.1 ng, while for Tnos in raw corn kernels was 0.01 ng. This in-house developed DNA extraction method is simple and obtains high-quality DNA suitable for GMO screening of 35S promotor and Tnos terminator in both raw and processed matrices.

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STUDIES ON A NON-MELATONIN PINEAL ANTI-GONADOTROPHIN

Acta Endocrinologica ◽

10.1530/acta.0.0690257 ◽

1972 ◽

Vol 69 (2) ◽

pp. 257-266 ◽

Cited By ~ 38

Author(s):

Bryant Benson ◽

Mary Jane Matthews ◽

Alvin E. Rodin

Keyword(s):

Solvent Extraction ◽

Organic Solvent ◽

Pineal Gland ◽

Gel Filtration ◽

Test System ◽

Pineal Body ◽

Physiological Effects ◽

Organic Solvent Extraction ◽

Gland Function

ABSTRACT Continuing investigation of pineal gland function indicates that the anti-gonadotrophic activity of this organ cannot be attributed solely to the postulated hormone melatonin, the concentration of which is negligible in the pineal body compared to quantities required to produce unequivocal physiological effects. A non-melatonin antigonadotrophic substance recently isolated from bovine pineal glands was further purified by organic solvent extraction, ultrafiltration and gel filtration. Studies of partial blockage of compensatory ovarian hypertrophy in unilaterally ovariectomized Charles River CD-1 mice indicated that this substance is significantly more potent than melatonin in this test system.

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Facile fractionation of bamboo hydrolysate and characterization of isolated lignin and lignin-carbohydrate complexes

Holzforschung ◽

10.1515/hf-2020-0040 ◽

2020 ◽

Vol 0 (0) ◽

Author(s):

Xiaodi Wang ◽

Yongchao Zhang ◽

Luyao Wang ◽

Xiaoju Wang ◽

Qingxi Hou ◽

...

Keyword(s):

Solvent Extraction ◽

Organic Solvent ◽

Extraction Process ◽

Water Soluble ◽

Organic Solvent Extraction ◽

Glycoside Linkage ◽

Potential Applications ◽

Tremendous Amount ◽

Main Components ◽

Hemicellulosic Sugars

AbstractAn efficient separation technology for hydrolysates towards a full valorization of bamboo is still a tough challenge, especially regarding the lignin and lignin-carbohydrate complexes (LCCs). The present study aimed to develop a facile approach using organic solvent extraction for efficiently fractionating the main components of bamboo hydrolysates. The high-purity lignin with only a trace of carbohydrates was first obtained by precipitation of the bamboo hydrolysate. The water-soluble lignin (WSL) fraction was extracted in organic solvent through a three-stage organic solvent extraction process, and the hemicellulosic sugars with increased purity were also collected. Furthermore, a thorough characterization including various NMR techniques (31P, 13C, and 2D-HSQC), GPC, and GC-MS was conducted to the obtained lignin-rich-fractions. It was found that the WSL fraction contained abundant functional groups and tremendous amount of LCC structures. As compared to native LCC of bamboo, the WSL fraction exhibited more typical LCC linkages, i.e. phenyl glycoside linkage, which is the main type of chemical linkage between lignin and carbohydrate in both LCC samples. The results demonstrate that organic phase extraction is a highly efficient protocol for the fractionation of hydrolysate and the isolation of LCC-rich streams possessing great potential applications.

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