A general method of isolating high molecular weight DNA from methanogenic archaea (archaebacteria)

1992 ◽  
Vol 38 (1) ◽  
pp. 65-68 ◽  
Author(s):  
Ken F. Jarrell ◽  
David Faguy ◽  
Anne M. Hebert ◽  
Martin L. Kalmokoff
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Thermal Denaturation ◽  
Dna Isolation ◽  
Methanogenic Archaea ◽  
Restriction Enzymes ◽  
Mole Percent ◽  
High Molecular Weight Dna ◽  
Isolation Methods ◽  
General Method

High molecular weight DNA was readily isolated from all methanogens treated, as well as from thermophilic anaerobic eubacteria, by grinding cells frozen in liquid N2, prior to lysis with SDS. DNA can subsequently be purified by the usual phenol–chloroform extractions. The procedure yields DNA readily cut by restriction enzymes and suitable for oligonucleotide probing, as well as for mole percent G + C content determination by thermal denaturation. The method routinely yields DNA of high molecular weight and is an improvement over DNA isolation methods for many methanogens, which often involve an initial breakage of the cells in a French pressure cell. Key words: methanogens, archaebacteria, archaea, DNA isolation.

NUTRITIONAL AND THERAPEUTIC EVALUATION OF SPIRULINA PLATENSIS

2020 ◽  
pp. 86-90
Author(s):  
SACHIN KULKARNI ◽  
DEEPALI CHAVAN
Keyword(s):  
Molecular Weight ◽  
Protein Content ◽  
High Molecular Weight ◽  
Spirulina Platensis ◽  
Dna Isolation ◽  
Protein Profile ◽  
Polyacrylamide Gel Electrophoresis ◽  
Growth Monitoring ◽  
High Molecular Weight Dna ◽  
Sds Page

Objective: The present investigation was to isolate and produce Spirulina platensis on high scale for food pharmaceutical and aquaculture due to the presence of high protein content. Methods: Cultivation of Spirulina preparation of culture medium, inoculums build up, growth monitoring, harvesting, drying, procedure for protein estimation, sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and DNA isolation of Spirulina platensis. Results: The study shows that protein content of S. platensis was found to be 62% protein profile was studied through SDS-PAGE, the bands were in the range of 35 kDa–44 kDa. S. platensis contain high molecular weight DNA. The bands was seen and visualize under the transilluminator it reveals that the alga S. platensis contain high molecular weight DNA. Conclusion: The production of Spirulina by simple pH determination method suggests economic production of alga by a simple process. The DNA isolation showing that these algae contain high molecular weight DNA. However, utilization of this biomass for varied end uses, such as food, feed, aquaculture, and pharmaceuticals.

scholarly journals High molecular weight DNA isolation method from diverse plant species for use with Oxford Nanopore sequencing

2019 ◽  
Author(s):  
Brieanne Vaillancourt ◽  
C. Robin Buell
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Dna Isolation ◽  
Isolation Method ◽  
High Molecular Weight Dna ◽  
Sequencing Platform ◽  
Long Reads ◽  
Oxford Nanopore ◽  
Long Read ◽  
Oxford Nanopore Technologies

AbstractThe ability to generate long reads on the Oxford Nanopore Technologies sequencing platform is dependent on the isolation of high molecular weight DNA free of impurities. For some taxa, this is relatively straightforward; however, for plants, the presence of cell walls and a diverse set of specialized metabolites such as lignin, phenolics, alkaloids, terpenes, and flavonoids present significant challenges in the generation of DNA suitable for production of long reads. Success in generating long read lengths and genome assemblies of plants has been reported using diverse DNA isolation methods, some of which were tailored to the target species and/or required extensive labor. To avoid the need to optimize DNA isolation for each species, we developed a taxa-independent DNA isolation method that is relatively simple and efficient. This method expands on the Oxford Nanopore Technologies high molecular weight genomic DNA protocol from plant leaves and utilizes a conventional cetyl trimethylammonium bromide extraction followed by removal of impurities and short DNA fragments using commercially available kits that yielded robust N50 read lengths and yield on Oxford Nanopore Technologies flow cells.

Comparison of two methods of high-molecular-weight DNA isolation from human leucocytes

1983 ◽  
Vol 134 (2) ◽  
pp. 320-324 ◽  
Author(s):  
Chantal Gautreau ◽  
Cécile Rahuel ◽  
Jean-Pierre Cartron ◽  
Gérard Lucotte
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Dna Isolation ◽  
High Molecular Weight Dna

scholarly journals Genomic DNA isolation from scrophularieae dried leaves using a simple, high-throughput protocol

2019 ◽  
Vol 48 (4) ◽  
pp. 1231-1235
Author(s):  
Mehrshid Riahi ◽  
Melina Babaei ◽  
Farrokh Ghahremaninejad
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Dna Extraction ◽  
High Throughput ◽  
Genomic Dna ◽  
Dna Isolation ◽  
High Molecular Weight Dna ◽  
Plant Dna ◽  
Genomic Dna Isolation

This communication described efficient DNA extraction from Scrophularia and Verbascum samples. Modified Murray and Thompson modified Cota-Sànchez method and Bioflux kit methods were applied for the extraction of DNA. Among the different methods, Bioflux kit Plant DNA extraction kit, coupled with some modification was the best for extraction of high molecular weight DNA as long as the extracted DNA is quantified with fluorescence-based methods.

Isolation of high molecular weight DNA from wholeEuglena cells

PROTOPLASMA ◽  
1987 ◽  
Vol 141 (2-3) ◽  
pp. 139-148 ◽  
Author(s):  
Christine Mederic ◽  
Odile Bertaux ◽  
J. D. Rouzeau ◽  
R. Valencia
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
High Molecular Weight Dna

Role of proteolysis in apoptosis: involvement of serine proteases in internucleosomal DNA fragmentation in immature thymocytes

1993 ◽  
Vol 71 (9-10) ◽  
pp. 488-500 ◽  
Author(s):  
Valerie M. Weaver ◽  
Boleslaw Lach ◽  
P. Roy Walker ◽  
Marianna Sikorska
Keyword(s):  
Molecular Weight ◽  
Serine Protease ◽  
High Molecular Weight ◽  
Protease Inhibitors ◽  
Dna Fragmentation ◽  
Dna Cleavage ◽  
Apoptotic Cell ◽  
Dependent Manner ◽  
Serine Protease Inhibitors ◽  
High Molecular Weight Dna

Three chemically distinct serine, but not cysteine, protease inhibitors (phenylmethylsulphonyl fluoride, N-tosyl-L-phenylalanylchloromethyl ketone and 3,4-dichloroisocoumarin) prevented, in a dose-dependent manner, the characteristic apoptotic internucleosomal DNA cleavage (DNA ladder) typically observed in thymocytes in response to dexamethasone and teniposide VM-26. This effect was not the result of a direct inhibition of the Ca2+, Mg2+-dependent endonuclease, since oligonucleosomal DNA cleavage occurred in the presence of these inhibitors in isolated nuclei. The proteolytic step occurred at a very early stage of apoptosis, and preincubation of thymocytes with the inhibitors before dexamethasone or teniposide VM-26 were added irreversibly suppressed ladder formation. This implied that the cellular effector(s) of these compounds preexisted and were not resynthesized in response to the inducers of apoptosis. Serine protease inhibitors also suppressed apoptotic cell shrinkage and complete nuclear collapse, suggesting that these morphological changes were directly related to internucleosomal fragmentation of DNA. However, the serine protease inhibitors did not prevent high molecular weight DNA cleavage (> 50 kilobases) that preceded the ladder formation and thymocytes still died by apoptosis. This supported the view that internucleosomal DNA cleavage, considered to be the biochemical marker of apoptosis, might in fact be a late and dispensable step and that the newly described high molecular weight DNA cleavage might be a better indicator of apoptosis.Key words: serine protease, apoptosis, internucleosomal DNA fragmentation, high molecular weight DNA cleavage, protease inhibitors.

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