scholarly journals Capture and elongation of single chromosomal DNA molecules using optically driven microchopsticks

2020 ◽  
Vol 14 (4) ◽  
pp. 044114
Author(s):  
Ryo Inukai ◽  
Hidekuni Takao ◽  
Fusao Shimokawa ◽  
Kyohei Terao
Keyword(s):  
Dna Molecules ◽  
Chromosomal Dna

scholarly journals An electrophoretic karyotype of Neurospora crassa.

1988 ◽  
Vol 8 (4) ◽  
pp. 1469-1473 ◽  
Author(s):  
M J Orbach ◽  
D Vollrath ◽  
R W Davis ◽  
C Yanofsky
Keyword(s):  
Neurospora Crassa ◽  
Electric Fields ◽  
Gel Electrophoresis ◽  
Electrophoretic Karyotype ◽  
Dna Molecules ◽  
Linkage Groups ◽  
Homogeneous Electric Field ◽  
Chromosomal Dna ◽  
Molecular Karyotype ◽  
Electrophoresis System

A molecular karyotype of Neurospora crassa was obtained by using an alternating-field gel electrophoresis system which employs contour-clamped homogeneous electric fields. The migration of all seven N. crassa chromosomal DNAs was defined, and five of the seven molecules were separated from one another. The estimated sizes of these molecules, based on their migration relative to Schizosaccharomyces pombe chromosomal DNA molecules, are 4 to 12.6 megabases. The seven linkage groups were correlated with specific chromosomal DNA bands by hybridizing transfers of contour-clamped homogeneous electric field gels with radioactive probes specific to each linkage group. The mobilities of minichromosomal DNAs generated from translocation strains were also examined. The methods used for preparation of chromosomal DNA molecules and the conditions for their separation should be applicable to other filamentous fungi.

scholarly journals Electrophoretic analysis of Histoplasma capsulatum chromosomal DNA.

1989 ◽  
Vol 9 (3) ◽  
pp. 983-987 ◽  
Author(s):  
P E Steele ◽  
G F Carle ◽  
G S Kobayashi ◽  
G Medoff
Keyword(s):  
Gel Electrophoresis ◽  
Histoplasma Capsulatum ◽  
Electrophoretic Analysis ◽  
Considerable Variability ◽  
Dna Molecules ◽  
Homogeneous Electric Field ◽  
Chromosomal Dna ◽  
Experimental Conditions ◽  
Agarose Gels ◽  
Minimum Estimate

Seven chromosome-sized DNA molecules in the Downs strain of Histoplasma capsulatum were resolved by using chromosome-specific DNA probes in blot hybridizations of contour-clamped homogeneous electric field (CHEF) and field-inversion gel electrophoresis (FIGE) agarose gels. The sizes of the chromosomal DNA bands extended from that of the largest Saccharomyces cerevisiae chromosome to beyond that of the Schizosaccharomyces pombe chromosomes. Under our experimental conditions, the order of the five largest DNA bands was inverted in the FIGE gel relative to the CHEF gel, demonstrating a characteristic of FIGE whereby large DNA molecules may have greater rather than lesser mobility with increasing size. Comparison of the Downs strain with other H. capsulatum strains by CHEF and FIGE analysis revealed considerable variability in band mobility. The resolution of seven chromosome-sized DNA molecules in the Downs strain provides a minimum estimate of the chromosome number.

The form of chromosomal DNA molecules in bacterial cells

Biochimie ◽  
2001 ◽  
Vol 83 (2) ◽  
pp. 177-186 ◽  
Author(s):  
Arnold J Bendich
Keyword(s):  
Bacterial Cells ◽  
Dna Molecules ◽  
Chromosomal Dna

Electrophoretic analysis of Histoplasma capsulatum chromosomal DNA

1989 ◽  
Vol 9 (3) ◽  
pp. 983-987
Author(s):  
P E Steele ◽  
G F Carle ◽  
G S Kobayashi ◽  
G Medoff
Keyword(s):  
Gel Electrophoresis ◽  
Histoplasma Capsulatum ◽  
Electrophoretic Analysis ◽  
Considerable Variability ◽  
Dna Molecules ◽  
Homogeneous Electric Field ◽  
Chromosomal Dna ◽  
Experimental Conditions ◽  
Agarose Gels ◽  
Minimum Estimate

Seven chromosome-sized DNA molecules in the Downs strain of Histoplasma capsulatum were resolved by using chromosome-specific DNA probes in blot hybridizations of contour-clamped homogeneous electric field (CHEF) and field-inversion gel electrophoresis (FIGE) agarose gels. The sizes of the chromosomal DNA bands extended from that of the largest Saccharomyces cerevisiae chromosome to beyond that of the Schizosaccharomyces pombe chromosomes. Under our experimental conditions, the order of the five largest DNA bands was inverted in the FIGE gel relative to the CHEF gel, demonstrating a characteristic of FIGE whereby large DNA molecules may have greater rather than lesser mobility with increasing size. Comparison of the Downs strain with other H. capsulatum strains by CHEF and FIGE analysis revealed considerable variability in band mobility. The resolution of seven chromosome-sized DNA molecules in the Downs strain provides a minimum estimate of the chromosome number.

scholarly journals Development of gene mapping for single chromosomal DNA molecules

1999 ◽  
Vol 119 (12) ◽  
pp. 620-625 ◽  
Author(s):  
Yasunori Matsui ◽  
Kazunori Takashima ◽  
Shinji Katsura ◽  
Akira Mizuno
Keyword(s):  
Gene Mapping ◽  
Dna Molecules ◽  
Chromosomal Dna

scholarly journals Packaging specific segments of the Salmonella chromosome with locked-in Mud-P22 prophages.

Genetics ◽  
1988 ◽  
Vol 118 (4) ◽  
pp. 581-592
Author(s):  
P Youderian ◽  
P Sugiono ◽  
K L Brewer ◽  
N P Higgins ◽  
T Elliott
Keyword(s):  
Homologous Recombination ◽  
Genetic Markers ◽  
Dna Molecules ◽  
Chromosomal Dna ◽  
Genetic Elements ◽  
Phage P22 ◽  
Salmonella Phage

Abstract Hybrid genetic elements, Mud-P and Mud-Q (collectively, Mud-P22s), have been constructed that carry two-thirds of the temperate Salmonella phage P22 genome sandwiched between the ends of transposon Mu. Insertions of these elements in the Salmonella chromosome generate locked-in P22 prophages that cannot excise. Upon induction (as a consequence of the inactivation of P22 c2 repressor), a locked-in prophage replicates its DNA in situ, resulting in the amplification of neighboring regions of the chromosome and the processive packaging of three contiguous headsful of adjacent DNA in one direction from the P22 packaging site, pac. Phage particles in an induced lysate of a Mud-P22 lysogen contain DNA molecules corresponding to several minutes of chromosomal DNA adjacent to the site of prophage insertion and transduce nearby genetic markers with high efficiencies. Mud-P22 prophages have been introduced into an F' episome by transposition; resident Mud insertions on the Salmonella chromosome may be converted to Mud-P22 insertions by homologous recombination in P22-mediated transductional crosses.

scholarly journals On-Site Processing of Single Chromosomal DNA Molecules Using Optically Driven Microtools on A Microfluidic Workbench

2020 ◽  
Author(s):  
Akihito Masuda ◽  
Hidekuni Takao ◽  
Fusao Shimokawa ◽  
KYOHEI TERAO
Keyword(s):  
Optical Tweezers ◽  
Single Molecule ◽  
Single Cells ◽  
Optical Microscope ◽  
Optical Manipulation ◽  
Confined Space ◽  
Dna Molecules ◽  
Chromosomal Dna ◽  
Single Molecule Level ◽  
Immobilization Of Enzymes

Abstract We developed optically driven microtools for processing single biomolecules using a microfluidic workbench composed of a microfluidic platform that functions under an optical microscope. The optically driven microtools have enzymes immobilized on their surfaces, which catalyze chemical reactions for molecular processing in a confined space. Optical manipulation of the microtools enables them to be integrated with a microfluidic device for controlling the position, orientation, shape of the target sample. Here, we describe the immobilization of enzymes on the surface of microtools, the microfluidics workbench, including its microtool storage and sample positioning functions, and the use of this system for on-site cutting of single chromosomal DNA molecules. We fabricated microtools by UV lithography with SU-8 and selected ozone treatments for immobilizing enzymes. The microfluidic workbench has tool-stock chambers for tool storage and micropillars to trap and extend single chromosomal DNA molecules. The DNA cutting enzymes DNaseI and DNaseII were immobilized on microtools that were manipulated using optical tweezers. The DNaseI tool shows reliable cutting for on-site processing. This pinpoint processing provides an approach for analyzing chromosomal DNA at the single-molecule level. The flexibility of the microtool design allows for processing of various samples, including biomolecules and single cells.

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