Planarian High Molecular Weight DNA Isolation by Spooling

Author(s):  
Shasha Zhang ◽  
Alejandro Sánchez Alvarado
1992 ◽  
Vol 38 (1) ◽  
pp. 65-68 ◽  
Author(s):  
Ken F. Jarrell ◽  
David Faguy ◽  
Anne M. Hebert ◽  
Martin L. Kalmokoff

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.


Author(s):  
SACHIN KULKARNI ◽  
DEEPALI CHAVAN

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.


2019 ◽  
Author(s):  
Brieanne Vaillancourt ◽  
C. Robin Buell

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.


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

2019 ◽  
Vol 48 (4) ◽  
pp. 1231-1235
Author(s):  
Mehrshid Riahi ◽  
Melina Babaei ◽  
Farrokh Ghahremaninejad

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.


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

1993 ◽  
Vol 71 (9-10) ◽  
pp. 488-500 ◽  
Author(s):  
Valerie M. Weaver ◽  
Boleslaw Lach ◽  
P. Roy Walker ◽  
Marianna Sikorska

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.


2019 ◽  
Author(s):  
Rasmus Dam Wollenberg ◽  
Brian Strehlow ◽  
Astrid Schuster

This protocol was used to extract high molecular weight DNA from the sponge (porifera) Halichondria panicea. This protocol contains slight modifications from that presented in Ausubel et al (1995). References: Ausubel, F.; Brent, R.; Kingston, R.; Moore, D.; Seidman, J.G.; Smith, J.;Struhl, K. Short Protocols in Molecular Biology(1995), 3rd ed., Unit 2.1: page 2-3


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