Abstract
A number of recent developments in DNA analysis by capillary electrophoresis are here reviewed. They include capillary arrays for fast, parallel DNA sequencing as well as microfabricated capillary arrays. Microfluidic chips for DNA sizing and quantitation are also covered, as well as microdevices containing arrays of regular obstacles acting as size-separators during DNA migration. Screening of DNA point mutations by two much improved techniques is also reported: in one case, such mutations are detected (but only on relative short, ca. 60-70 base-long fragments) by free electrophoresis in rather acidic (pH ca. 3) buffers; in the case of single-strand chain polymorphism, an improved technique is described based on near-neutral pH buffers with mixtures of Tris/MES cations/zwitterions. When studying the behavior of inorganic and organic cations in the Debye-Hückel layer of DNA, it was found that the latter (especially a large number of Good's buffers and other zwitterions, such as His) would bind to the DNA filament not only via charge interaction, but also via additional bonds, notably hydrogen bonds, thus altering the electrophoretic (and possibly the biological) behavior of DNA molecules. However, whether or not borate ions would bind to DNA remains still an unsettled question. Finally, capillary electrophoresis was found to be instrumental in measuring fine physicochemical parameters pertaining to DNA polyelectrolytes, such as their free mobility and their translational diffusion coefficients.
| Original language | English |
|---|---|
| Pages (from-to) | 1361-1374 |
| Number of pages | 14 |
| Journal | Electrophoresis |
| Volume | 23 |
| Issue number | 10 |
| DOIs | |
| Publication status | Published - 2002 |
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Keywords
- DNA separations
- DNA sequencing
- Microchip devices
- Point mutations
- Review
ASJC Scopus subject areas
- Clinical Biochemistry
Cite this
Recent progress in DNA analysis by capillary electrophoresis. / Righetti, Pier Giorgio; Gelfi, Cecilia; D'Acunto, Maria Rosa.
In: Electrophoresis, Vol. 23, No. 10, 2002, p. 1361-1374.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Recent progress in DNA analysis by capillary electrophoresis
AU - Righetti, Pier Giorgio
AU - Gelfi, Cecilia
AU - D'Acunto, Maria Rosa
PY - 2002
Y1 - 2002
N2 - A number of recent developments in DNA analysis by capillary electrophoresis are here reviewed. They include capillary arrays for fast, parallel DNA sequencing as well as microfabricated capillary arrays. Microfluidic chips for DNA sizing and quantitation are also covered, as well as microdevices containing arrays of regular obstacles acting as size-separators during DNA migration. Screening of DNA point mutations by two much improved techniques is also reported: in one case, such mutations are detected (but only on relative short, ca. 60-70 base-long fragments) by free electrophoresis in rather acidic (pH ca. 3) buffers; in the case of single-strand chain polymorphism, an improved technique is described based on near-neutral pH buffers with mixtures of Tris/MES cations/zwitterions. When studying the behavior of inorganic and organic cations in the Debye-Hückel layer of DNA, it was found that the latter (especially a large number of Good's buffers and other zwitterions, such as His) would bind to the DNA filament not only via charge interaction, but also via additional bonds, notably hydrogen bonds, thus altering the electrophoretic (and possibly the biological) behavior of DNA molecules. However, whether or not borate ions would bind to DNA remains still an unsettled question. Finally, capillary electrophoresis was found to be instrumental in measuring fine physicochemical parameters pertaining to DNA polyelectrolytes, such as their free mobility and their translational diffusion coefficients.
AB - A number of recent developments in DNA analysis by capillary electrophoresis are here reviewed. They include capillary arrays for fast, parallel DNA sequencing as well as microfabricated capillary arrays. Microfluidic chips for DNA sizing and quantitation are also covered, as well as microdevices containing arrays of regular obstacles acting as size-separators during DNA migration. Screening of DNA point mutations by two much improved techniques is also reported: in one case, such mutations are detected (but only on relative short, ca. 60-70 base-long fragments) by free electrophoresis in rather acidic (pH ca. 3) buffers; in the case of single-strand chain polymorphism, an improved technique is described based on near-neutral pH buffers with mixtures of Tris/MES cations/zwitterions. When studying the behavior of inorganic and organic cations in the Debye-Hückel layer of DNA, it was found that the latter (especially a large number of Good's buffers and other zwitterions, such as His) would bind to the DNA filament not only via charge interaction, but also via additional bonds, notably hydrogen bonds, thus altering the electrophoretic (and possibly the biological) behavior of DNA molecules. However, whether or not borate ions would bind to DNA remains still an unsettled question. Finally, capillary electrophoresis was found to be instrumental in measuring fine physicochemical parameters pertaining to DNA polyelectrolytes, such as their free mobility and their translational diffusion coefficients.
KW - DNA separations
KW - DNA sequencing
KW - Microchip devices
KW - Point mutations
KW - Review
UR - http://www.scopus.com/inward/record.url?scp=0036078673&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0036078673&partnerID=8YFLogxK
U2 - 10.1002/1522-2683(200205)23:10<1361::AID-ELPS1361>3.0.CO;2-J
DO - 10.1002/1522-2683(200205)23:10<1361::AID-ELPS1361>3.0.CO;2-J
M3 - Article
C2 - 12116146
AN - SCOPUS:0036078673
VL - 23
SP - 1361
EP - 1374
JO - Electrophoresis
JF - Electrophoresis
SN - 0173-0835
IS - 10
ER -