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Title: High-Throughput Genetic Analysis and Combinatorial Chiral Separations Based on Capillary Electrophoresis

Abstract

Capillary electrophoresis (CE) offers many advantages over conventional analytical methods, such as speed, simplicity, high resolution, low cost, and small sample consumption, especially for the separation of enantiomers. However, chiral method developments still can be time consuming and tedious. They designed a comprehensive enantioseparation protocol employing neutral and sulfated cyclodextrins as chiral selectors for common basic, neutral, and acidic compounds with a 96-capillary array system. By using only four judiciously chosen separation buffers, successful enantioseparations were achieved for 49 out of 54 test compounds spanning a large variety of pKs and structures. Therefore, unknown compounds can be screened in this manner to identify optimal enantioselective conditions in just one rn. In addition to superior separation efficiency for small molecules, CE is also the most powerful technique for DNA separations. Using the same multiplexed capillary system with UV absorption detection, the sequence of a short DNA template can be acquired without any dye-labels. Two internal standards were utilized to adjust the migration time variations among capillaries, so that the four electropherograms for the A, T, C, G Sanger reactions can be aligned and base calling can be completed with a high level of confidence. the CE separation of DNA can bemore » applied to study differential gene expression as well. Combined with pattern recognition techniques, small variations among electropherograms obtained by the separation of cDNA fragments produced from the total RNA samples of different human tissues can be revealed. These variations reflect the differences in total RNA expression among tissues. Thus, this Ce-based approach can serve as an alternative to the DNA array techniques in gene expression analysis.« less

Authors:
 [1]
  1. Iowa State Univ., Ames, IA (United States)
Publication Date:
Research Org.:
Ames Lab., Ames, IA (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
816440
Report Number(s):
IS-T 2004
TRN: US200320%%544
DOE Contract Number:  
W-7405-Eng-82
Resource Type:
Thesis/Dissertation
Resource Relation:
Other Information: TH: Thesis (Ph.D.); Submitted to Iowa State Univ., Ames, IA (US); PBD: 5 Aug 2003
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; ABSORPTION; ANIMAL TISSUES; BUFFERS; CAPILLARIES; DETECTION; DNA; EFFICIENCY; ELECTROPHORESIS; ENANTIOMORPHS; GENES; GENETICS; PATTERN RECOGNITION; RESOLUTION; RNA; VELOCITY

Citation Formats

Zhong, Wenwan. High-Throughput Genetic Analysis and Combinatorial Chiral Separations Based on Capillary Electrophoresis. United States: N. p., 2003. Web. doi:10.2172/816440.
Zhong, Wenwan. High-Throughput Genetic Analysis and Combinatorial Chiral Separations Based on Capillary Electrophoresis. United States. https://doi.org/10.2172/816440
Zhong, Wenwan. Wed . "High-Throughput Genetic Analysis and Combinatorial Chiral Separations Based on Capillary Electrophoresis". United States. https://doi.org/10.2172/816440. https://www.osti.gov/servlets/purl/816440.
@article{osti_816440,
title = {High-Throughput Genetic Analysis and Combinatorial Chiral Separations Based on Capillary Electrophoresis},
author = {Zhong, Wenwan},
abstractNote = {Capillary electrophoresis (CE) offers many advantages over conventional analytical methods, such as speed, simplicity, high resolution, low cost, and small sample consumption, especially for the separation of enantiomers. However, chiral method developments still can be time consuming and tedious. They designed a comprehensive enantioseparation protocol employing neutral and sulfated cyclodextrins as chiral selectors for common basic, neutral, and acidic compounds with a 96-capillary array system. By using only four judiciously chosen separation buffers, successful enantioseparations were achieved for 49 out of 54 test compounds spanning a large variety of pKs and structures. Therefore, unknown compounds can be screened in this manner to identify optimal enantioselective conditions in just one rn. In addition to superior separation efficiency for small molecules, CE is also the most powerful technique for DNA separations. Using the same multiplexed capillary system with UV absorption detection, the sequence of a short DNA template can be acquired without any dye-labels. Two internal standards were utilized to adjust the migration time variations among capillaries, so that the four electropherograms for the A, T, C, G Sanger reactions can be aligned and base calling can be completed with a high level of confidence. the CE separation of DNA can be applied to study differential gene expression as well. Combined with pattern recognition techniques, small variations among electropherograms obtained by the separation of cDNA fragments produced from the total RNA samples of different human tissues can be revealed. These variations reflect the differences in total RNA expression among tissues. Thus, this Ce-based approach can serve as an alternative to the DNA array techniques in gene expression analysis.},
doi = {10.2172/816440},
url = {https://www.osti.gov/biblio/816440}, journal = {},
number = ,
volume = ,
place = {United States},
year = {2003},
month = {1}
}

Thesis/Dissertation:
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