scholarly journals The Implications of Fragmented Genomic DNA Size Range on the Hybridization Efficiency in NanoGene Assay

Sensors ◽  
2018 ◽  
Vol 18 (8) ◽  
pp. 2646 ◽  
Author(s):  
Xiaofang Wang ◽  
Beelee Chua ◽  
Ahjeong Son

DNA hybridization-based assays are well known for their ability to detect and quantify specific bacteria. Assays that employ DNA hybridization include a NanoGene assay, fluorescence in situ hybridization, and microarrays. Involved in DNA hybridization, fragmentation of genomic DNA (gDNA) is necessary to increase the accessibility of the probe DNA to the target gDNA. However, there has been no thorough and systematic characterization of different fragmented gDNA sizes and their effects on hybridization efficiency. An optimum fragmented size range of gDNA for the NanoGene assay is hypothesized in this study. Bacterial gDNA is fragmented via sonication into different size ranges prior to the NanoGene assay. The optimum size range of gDNA is determined via the comparison of respective hybridization efficiencies (in the form of quantification capabilities). Different incubation durations are also investigated. Finally, the quantification capability of the fragmented (at optimum size range) and unfragmented gDNA is compared.

Genome ◽  
1994 ◽  
Vol 37 (3) ◽  
pp. 477-481 ◽  
Author(s):  
Jie Xu ◽  
R. L. Conner ◽  
A. Laroche

'Agrotana', a wheat-alien hybrid (2n = 56), is a potential source of resistance to common root rot, stem rust, wheat streak mosaic virus, and the wheat curl mite. However, the origin of 'Agrotana', reported to be durum wheat × Agropyron trichophorum (pubescent wheatgrass), is uncertain. The objective of this investigation was to determine the chromosome constitution of 'Agrotana' using C-banding and fluorescence in situ hybridization techniques. The F1 hybrid of 'Agrotana' × 'Chinese Spring' wheat showed 7 I + 21 II in 14.9% of the pollen mother cells, evidence of the presence of the A, B, and D genomes in 'Agrotana'. The hybrid had 16 heavily C-banded chromosomes, namely 4A, and 1-7B of wheat, and a translocation that probably involved wheat chromosomes 2A and 2D. In situ hybridization using biotinylated genomic DNA of Ag. trichophorum cv. Greenleaf blocked with CS DNA failed to identify the alien chromosomes in 'Agrotana', indicating that the alien chromosomes were not likely derived from pubescent wheatgrass. In situ hybridization using labelled wheat genomic DNA blocked with 'Agrotana' DNA revealed that 'Agrotana' had 40 wheat, 14 alien, and 2 (a pair) wheat–alien translocated chromosomes. There was no homology between wheat and the alien chromosomes or chromosome segments involved in the wheat–alien recombinant. Two of the seven pairs of alien chromosomes were homoeologous to each other. The ability to identify alien chromatin in wheat using labelled wheat DNA instead of labelled alien DNA will be particularly useful in chromosome engineering of wheat germplasms having alien chromatin of unknown origin.Key words: wheat–alien hybrid, C-banding, fluorescence in situ hybridization, labelled wheat DNA as probe.


Genomics ◽  
1993 ◽  
Vol 17 (2) ◽  
pp. 403-407 ◽  
Author(s):  
Elizabeth A. Lindsay ◽  
Stephanie Halford ◽  
Roy Wadey ◽  
Peter J. Scambler ◽  
Antonio Baldini

Author(s):  
Oliver A. Press ◽  
Roberta Guzman ◽  
Monica Cervantes ◽  
Angela Santiago ◽  
Michael F. Press

2008 ◽  
Vol 182 (2) ◽  
pp. 116-121 ◽  
Author(s):  
Zsuzsa Rákosy ◽  
Laura Vízkeleti ◽  
Szilvia Ecsedi ◽  
Ágnes Bégány ◽  
Gabriella Emri ◽  
...  

1993 ◽  
Vol 47 (8) ◽  
pp. 1153-1156 ◽  
Author(s):  
Alessandra M. V. Duncan ◽  
Athen Macdonald ◽  
Carolyn J. Brown ◽  
Daynna Wolff ◽  
Huntington F. Willard ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document