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Evaluation of sieving matrices used to separate alleles by cycling temperature capillary electrophoresis
Author(s) -
Ekstrøm Per Olaf,
Bjørheim Jens
Publication year - 2006
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500642
Subject(s) - capillary electrophoresis , matrix (chemical analysis) , chromatography , polymer , reproducibility , acrylamide , polyacrylamide , dissolution , allele , consumables , analytical chemistry (journal) , chemistry , materials science , polymer chemistry , biochemistry , monomer , organic chemistry , gene
Denaturing CE (DCE) is a powerful tool for analysis of DNA variation. The development of commercial multi‐CE instruments allows large‐scale studies of DNA variation (many samples and many fragments). However, the cost of consumables like capillary arrays and sieving matrix might limit the use of DCE in such studies. Thus, we have tested 72 different in‐house formulated sieving matrices' ability to suppress EOF and separate PCR‐amplified alleles with the DCE variant, cycling temperature CE (CTCE). The data herein demonstrate that alleles can be baseline‐separated by use of PVP and poly( N , N ‐dimethyl acrylamide) polymers at various percentages and pH. Allele separation by CTCE is matrix‐independent and consequently applicable to any capillary instrument used for DNA separation. Formulation of sieving matrix for CTCE was done by dissolving appropriate amount of polymer powder into the running buffers. Allele separation was observed at different pH (7.5–8.5), concentrations and molecular size of the polymer, without compromising the separation and reproducibility. Finally, the cost reduction of homemade matrices is more than 1000‐fold as compared to commercial sieving matrices.