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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
14
pubmed:dateCreated
2002-9-4
pubmed:abstractText
The stepwise gradient of linear polymer matrices in microchannel electrophoresis is proposed as a means of achieving high-resolution separation of DNA samples containing a wide range of fragment sizes. In this method, multiple discrete steps in terms of polymer type or concentration are created in the microchannel by injecting appropriate solutions in order. The mixing of the various steps is found to be negligible compared to the effective length of separation channel, confirming that a stepwise gradient of matrices is formed. This technique is successfully applied to the analysis of restriction digest fragments and DNA ladders, and is demonstrated to provide higher resolution than the isocratic method, for both small and large fragments simultaneously. Even though the stepwise gradient is created manually, the reproducibility of the migration times of fragments in DNA samples is found to be quite good. Taken the separation of 100 bp DNA ladder in three steps gradient pattern as an example, the relative standard deviations of migration times are respectively less than 0.53% and 3.1% in six consecutive injections in one channel and in different channels. The migration of DNA fragments in gradient mode is shown to be similar to that for the isocratic scheme, allowing the design of each step to be made in reference to existing knowledge. These promising results indicate the great potential of this stepwise gradient method for the analysis of DNA by microchip electrophoresis, offering both high resolution and good reproducibility.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0173-0835
pubmed:author
pubmed:issnType
Print
pubmed:volume
23
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
2341-6
pubmed:meshHeading
pubmed:year
2002
pubmed:articleTitle
Stepwise gradient of linear polymer matrices in microchip electrophoresis for high-resolution separation of DNA.
pubmed:affiliation
Department of Medicinal Chemistry, Faculty of Pharmaceutical Sciences, The University of Tokushima, Japan Science and Technology Corporation (JST), Shomachi, Tokushima, Japan. lzhang@ph.tokushima-u.ac.jp
pubmed:publicationType
Journal Article