Comparison of DNA Extraction Methods in Terms of Yield, Purity, Long-Term Storage, and Downstream Manipulation with Brewer's Yeast Chromosomal DNA

Author(s):  
Kopecka
Zootaxa ◽  
2021 ◽  
Vol 4950 (1) ◽  
pp. 181-188
Author(s):  
ROXANA TIMM ◽  
MARTIN SCHWENTNER ◽  
SIMON BOBER ◽  
ANNE-NINA LÖRZ

Intact voucher specimens are essential to allow detailed morphological observations on specimens that are used in molecular genetic studies. This can be achieved either by dissection of small, taxonomically uninformative parts of the body for DNA extraction or by employing non-destructive DNA extraction methods. The latter is particularly important for small-bodied animals. Here we test the effects of Chelex-based DNA extraction on the integrity of setae and setules in Amphipoda, fragile structures of great taxonomic importance. Our results show that DNA extraction using Chelex had no influence on the setae and setule structure and is well suited for reverse taxonomic approaches and the long-term storage of morphological vouchers. A detailed protocol for non-destructive DNA extraction is provided. 


2019 ◽  
Author(s):  
Sudeshna Chakraborty ◽  
Anwesha Saha ◽  
N.A. Aravind

AbstractIsolation of high molecular weight DNA from gastropod molluscs and its subsequent PCR amplification is considered difficult due to excessive mucopolysaccharides secretion which co-precipitate with DNA and obstruct successful amplification. In an attempt to address this issue, we describe a modified CTAB DNA extraction method that proved to work significantly better with a number of freshwater and terrestrial gastropod taxa. We compared the performance of this method with Qiagen® DNeasy Blood and Tissue Kit. Reproducibility of amplification was verified using a set of taxon-specific primers wherein, modified CTAB extracted DNA could be replicated at least four out of five times but kit extracted DNA could not be replicated. Additionally, sequence quality was significantly better with CTAB extracted DNA. This could be attributed to the removal of polyphenolic compounds by polyvinyl pyrrolidone (PVP) which is the only difference between conventional and modified CTAB DNA extraction methods for animals. The genomic DNA isolated using modified CTAB protocol was of high quality (A260/280 ≥ 1.80) and could be used for downstream reactions even after long term storage (more than two years).


2001 ◽  
Vol 67 (4) ◽  
pp. 1682-1688 ◽  
Author(s):  
S. J. Williamson ◽  
M. R. McLaughlin ◽  
J. H. Paul

ABSTRACT The marine phage ΦHSIC has been previously reported to enter into a lysogenic relationship with its host, HSIC, identified asListonella pelagia. This phage produces a variety of plaques on its host, including turbid and haloed plaques, from which lysogens were previously isolated. These lysogens were unstable during long-term storage at −80° C and were lost. When HSIC was reinfected with phage ΦHSIC, pseudolysogen-like interactions between the phage and its host were observed. The cells (termed HSIC-2 or HSIC-2e) produced high viral titers (1011ml−1) in the absence of inoculating phage and yet reached culture densities of nearly 109 ml−1. Prophages were not induced by mitomycin C or the polyaromatic hydrocarbon naphthalene in cells harboring such infections. However, such cells were homoimmune to superinfection. Colonies hybridized strongly with a gene probe from a 100-bp fragment of the ΦHSIC genome, while the host did not. Analysis of chromosomal DNA preparations suggested the presence of a chromosomally integrated prophage. Phage adsorption experiments suggested that HSIC-2 was adsorption impaired. Because of the chromosomal prophage integration and homoimmunity, we interpret these results to indicate that ΦHSIC establishes a lysogenic relationship with its host that involves an extremely high level of spontaneous induction. This could be caused by a weak repressor of phage production. Additionally, poor phage adsorption of HSIC-2 compared to the wild type probably helped maintain this pseudolysogen-like relationship. In many ways, pseudolysogenic phage-host interactions may provide a paradigm for phage-host interactions in the marine environment.


2001 ◽  
Vol 6 (2) ◽  
pp. 3-14 ◽  
Author(s):  
R. Baronas ◽  
F. Ivanauskas ◽  
I. Juodeikienė ◽  
A. Kajalavičius

A model of moisture movement in wood is presented in this paper in a two-dimensional-in-space formulation. The finite-difference technique has been used in order to obtain the solution of the problem. The model was applied to predict the moisture content in sawn boards from pine during long term storage under outdoor climatic conditions. The satisfactory agreement between the numerical solution and experimental data was obtained.


Diabetes ◽  
1997 ◽  
Vol 46 (3) ◽  
pp. 519-523 ◽  
Author(s):  
G. M. Beattie ◽  
J. H. Crowe ◽  
A. D. Lopez ◽  
V. Cirulli ◽  
C. Ricordi ◽  
...  

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