scholarly journals Optimization of DNA extraction and amplification of Kikuyu (Pennisetum clandestinum Hochst. ex Chiov) for molecular identification

2021 ◽  
Vol 902 (1) ◽  
pp. 012016
Author(s):  
W Nawfetrias ◽  
J I Royani ◽  
I S Bidara ◽  
DP Handayani ◽  
M Surahman ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Molecular Identification ◽  
Crude Protein ◽  
Specific Primers ◽  
Sample Weight ◽  
Pennisetum Clandestinum ◽  
Annealing Temperatures ◽  
Optimized Protocol ◽  
High Yields ◽  
Pcr Conditions

Abstract Kikuyu (Pennisetum clandestinum Hochst. ex Chiov) is an important forage containing high crude protein for livestock. Molecular analysis of kikuyu relies on high yields of pure DNA and suitable PCR conditions. This research aimed to extract DNA from kikuyu based on weight of the sample and amplify the DNA of Burangrang accession using specific primers. 100 grams and 200 grams leaves of 3 accessions of kikuyu from Burangrang, Bukit Tunggul, and Tangkuban Perahu were extracted by Qiagen Mini Kit Plant. Concentration and purity of DNA were analyzed by NanoDrop Spectrophotometer 2000. DNA from Burangrang accession was amplified using six specific primers at different annealing temperatures. The result showed that the yield of DNA ranged 2.2 µg/µl to 21.4 µg/µl and the purity (ratio) were 1.08 to 2.01. Bukit Tunggul and Burangrang accession showed the same interaction pattern on the sample weight for concentration and purity. One hundred grams of leaves from Burangrang accession produce the highest concentration and the best purity of DNA, but no difference between other weight and accession. Reproducible amplifiable products were observed in all PCR reactions except primer K2. These results indicated that optimized protocol is suitable for further work on molecular identification of kikuyu.

scholarly journals Optimized Protocol for Cyanobacterial 16S rRNA Analysis in Danube Delta Lakes

2021 ◽  
Author(s):  
Maria Iasmina Moza ◽  
Carmen Postolache
Keyword(s):  
16S Rrna ◽  
Dna Extraction ◽  
Phytoplankton Biomass ◽  
Extraction Method ◽  
Seasonal Difference ◽  
Danube Delta ◽  
16S Rrna Analysis ◽  
Dna Extraction Method ◽  
Optimized Protocol ◽  
Cyanobacterial Biomass

AbstractMolecular biology protocols have been more and more accessible to researchers for ecological investigations, however, these protocols always require optimization steps for the analysis of specific types of samples. The purpose of this study was to optimize a molecular protocol for the analysis of cyanobacterial 16S rRNA in Danube Delta shallows lakes. In this regard, several commercial DNA extraction kits were tested in comparison with potassium ethyl xanthogenate extraction method on different matrices. The obtained DNA was further used for 16S rRNA PCR optimization. Finally, an optimized protocol is proposed for the molecular analysis of cyanobacteria group in freshwater samples. The best DNA extraction method was the potassium xanthogenate extraction from dried cyanobacterial biomass. A dynamic in total genomic eDNA was observed, reflecting the seasonal difference in phytoplankton biomass from the studied lakes. The PCR protocol optimized by us can be successfully applied for the identification of a broad range of cyanobacterial genetic markers.

Effects of Time of Nitrogen Application on Yield of Maize in the Tropics

1966 ◽  
Vol 2 (2) ◽  
pp. 101-105 ◽  
Author(s):  
Adeboyejo A. Fayemi
Keyword(s):  
Nitrogen Fertilizer ◽  
Crude Protein ◽  
Crude Protein Content ◽  
Nitrogen Application ◽  
Time Of Application ◽  
Cropping Season ◽  
Maize Grain ◽  
High Yields ◽  
The Tropics ◽  
Maize Ear

SummaryA four-year study from 1958 to 1962 showed that time of application of fertilizer nitrogen greatly influenced the yield of grain, the percentage of nitrogen and the crude protein of the grain under Nigerian conditions characteristic of the early maize cropping season from March to July. Split applications of nitrogen fertilizer significantly increased maize grain yield by 35 per cent when two equal doses were given one month and two months after planting; and by 31 per cent when four equal doses were supplied at planting and one month, two months and three months after seeding. Yield was significantly reduced when application was delayed two months after planting. High yields of maize were not obtained by supplying the whole of nitrogen fertilizer at one time, eidier at sowing or any time later during the growing season. However, applying all of the nitrogen fertilizer one month after planting significantly increased the percentage of nitrogen and of the crude protein content of the grain. The maize ear weight was favourably influenced by spreading the nitrogen application over the three-month period of the maize growth.

Rapid molecular detection ofEsteya vermicolabased on specific primers and the FTA-DNA extraction method

2014 ◽  
Vol 24 (8) ◽  
pp. 872-881 ◽  
Author(s):  
Ke Wei ◽  
Qinghua Wang ◽  
Yuzhu Wang ◽  
Liangjian Qu ◽  
Yong-an Zhang
Keyword(s):  
Dna Extraction ◽  
Extraction Method ◽  
Molecular Detection ◽  
Specific Primers ◽  
Dna Extraction Method

scholarly journals Evaluation and Application of an Efficient Plant DNA Extraction Protocol in Laboratory and Field Testing

2020 ◽  
Author(s):  
Qi Wang ◽  
Xiaoxia Shen ◽  
Tian Qiu ◽  
Wei Wu ◽  
Zhian Wang ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Molecular Identification ◽  
Filter Paper ◽  
Biological Samples ◽  
Gc Content ◽  
Dna Amplification ◽  
Nucleic Acid Amplification ◽  
Lamp Method ◽  
Paper Strip ◽  
Cellulose Filter

Abstract Background CTAB has been considered as the standard protocol for DNA extraction. But the complex and time-consuming procedures can’t meet the needs of rapid molecular identification. The method of using cellulose filter paper strips to transfer the DNA in the plant tissue lysate to the nucleic acid amplification system shortening the DNA extraction time to 30 s. However, cellulose filter paper strips have some shortcomings that cannot be put into widespread use. And the data supporting the rapidly purification of DNA by cellulose filter paper is not sufficient.ResultsIn the study, the published filter paper strip was modified by sticking the filter paper on the PVC sheet. This modified method is named EZ-D, for easy DNA extraction. Compared with the original method, the DNA extracted by EZ-D is more efficient in PCR amplification. We also came up with a new DNA extraction buffer, which exhibited higher DNA extraction efficiency. When compared with classic CTAB, EZ-D also showed great advantages for higher efficiency, easier protocol and lower cost. PCR analyses showed that DNA extracted from several types of plants by EZ-D were appropriate for specific identification of biological samples. PCR using DNA extracted by EZ-D was sensitive enough to detect 0.1 ng/μL. Evaluation of the EZ-D showed that the DNA extracts can be successfully amplified by PCR reaction for the DNA fragments up to 3000 bp in length and up to 80% in GC content. EZ-D was successfully used for DNA extraction from a variety of plant species and plant tissues. Moreover, when EZ-D was combined with the loop-mediated isothermal amplification (LAMP) method, DNA identification of biological samples could be achieved in an equipment-free way. Conclusion Combined with DNA amplification technology, EZ-D protocol is a rapid, specific and sensitive method for molecular identification of plant samples. In addition, EZ-D method is the first application of cellulose filter paper in the identification of genetically modified crops and traditional Chinese medicine ultra-fine powder. In terms of practicability, EZ-D has realized the popularization of cellulose filter paper for rapid DNA purification in laboratories and markets.

scholarly journals Yield and Nutritive Value of Sorghum, Maize and Soybean Forages Harvested in Southwestern Puerto Rico.

1969 ◽  
Vol 63 (3) ◽  
pp. 400-411
Author(s):  
D. G. St. Louis ◽  
J. A. Arroyo-Aguilú ◽  
A. Ramírez-Ortiz ◽  
R. E. McDowell
Keyword(s):  
Zea Mays ◽  
Puerto Rico ◽  
Glycine Max ◽  
Protein Content ◽  
Crude Protein ◽  
Nutritive Value ◽  
Grain Sorghum ◽  
Crude Protein Content ◽  
Forage Sorghum ◽  
High Yields

Six varieties of forage sorghum (Sorghum bicolor) and sorghum X sudangrass hybrids (S. bicolor X S. sudanese), six varieties of grain sorghum (S. bicolor) and three varieties each of maize (Zea mays) and soybean (Glycine max) were grown in 1.2 x 3.7 m plots at the Lajas Experiment Substation. All plots were harvested in the boot, flower and dough stages. Ratoons of the sorghum varieties were also harvested after 45 days of regrowth until the stands diminished. The forage sorghums and maize had higher yields of green and dry material than the grain sorghums. However, only the grain sorghums showed any significant increase in yield as the crop matured. All crops decreased in nutritive value with advance in age. In general, maize was higher in nutritive value than the forage sorghums. Results indicated that forage sorghums can be recommended due to high yields of good quality forage on irrigated lands on the south coast. However, results with soybean forage were poor. It does not appear feasible to produce soybeans to enhance the crude protein content of forage pellets.

scholarly journals Molecular diagnostics of bacterial and fungal plant diseases

2020 ◽  
Author(s):  
A. V. Sidarenka ◽  
H. A. Bareika ◽  
L. N. Valentovich ◽  
D. S. Paturemski ◽  
V. N. Kuptsou ◽  
...  
Keyword(s):  
Molecular Diagnostics ◽  
Plant Material ◽  
Plant Pathogens ◽  
Dna Isolation ◽  
Plant Diseases ◽  
Specific Primers ◽  
Fungal Plant Pathogens ◽  
Pcr Conditions ◽  
Soil And Water

Taxon-specific primers were developed and PCR conditions were optimized for diagnostics of bacterial and fungal plant pathogens. Methods for phytopathogens DNA isolation from plant material, soil and water were selected.

scholarly journals Evaluation of a PCR Primer Based on the Isocitrate Dehydrogenase Gene for Detection of Helicobacter pyloriin Feces

2000 ◽  
Vol 38 (10) ◽  
pp. 3755-3758 ◽  
Author(s):  
Frauke C. Argyros ◽  
Mayurika Ghosh ◽  
Lili Huang ◽  
Naoko Masubuchi ◽  
David R. Cave ◽  
...  
Keyword(s):  
Dna Extraction ◽  
Isocitrate Dehydrogenase ◽  
Specific Primers ◽  
Detection And Identification ◽  
Dna Base ◽  
Dehydrogenase Gene ◽  
H Pylori ◽  
Negative Controls ◽  
Lower Limits ◽  
Different Sources

In order to improve detection and identification ofHelicobacter pylori in highly contaminated samples, we evaluated new specific primers based on the DNA base sequence within the isocitrate dehydrogenase (icd) gene to amplify a 1,200-bp DNA segment. The specificity of the icd primer was tested against DNA derived from various bacteria, including 7Helicobacter species and a panel of 1 gram-variable, 2 gram-positive, and 16 gram-negative bacteria, as well as DNA from houseflies and feces from H. pylori-negative patients. The primers permitted the detection of all clinical H. pyloriisolates tested, but no reactions were observed with negative controls. Several procedures for DNA extraction from feces were evaluated using PCR with icd primers. The lower limits of detection ofH. pylori DNA from two different sources containing the same number of H. pylori organisms, a pure culture and feces spiked with H. pylori, were established for each extraction method tested. The results were 8.0 × 103CFU/ml for cultures of pure H. pylori, and 8.0 × 106 CFU/ml for H. pylori from feces, using the phenol-chloroform method; 8.0 × 102 and 7.0 × 103 CFU/ml, respectively, for a glass matrix and chaotropic solution protocol; 8.0 × 102 and 7.0 × 103 CFU/ml, respectively, for the QIAamp tissue kit; and 5.0 × 102 and 5.0 × 103 CFU/ml, respectively, for the XTRAX DNA extraction kit. We conclude that the use of the icd gene as a primer for PCR represents a specific and sensitive assay for detection of H. pylori in highly contaminated samples.

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