Separation of DNA by capillary electrophoresis.
This chapter reports an overview of the analytical techniques used to perform genetic analysis of polymerase chain reaction products using capillary electrophoresis. Three separate but related techniques are described: the separation of native DNA with detection using fluorescent intercalating dyes, the separation of denatured DNA using fluorescently labeled primers, and the detection of single-strand conformation polymorphisms using denatured DNA separated under native conditions. The various techniques involve electrokinetic injection of the DNA onto a narrow band at the head of the column, sieving the DNA through various entangled polymer matrices, and detection via single or multichannel laser-induced fluorescence. Analytical protocols are provided, and a series of representative electropherograms are included.
McCord
Bruce
B
Department of Chemistry, Florida International University, University Park, Miami, FL, USA.
Hartzell-Baguley
Brittany
B
King
Stephanie
S
D004247:DNA / Q000302:isolation & purification*; D004275:DNA, Ribosomal / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000379:methods* / Q000592:standards; D006801:Humans; D018895:Microsatellite Repeats / Q000235:genetics; D009691:Nucleic Acid Denaturation; D010957:Plasmids / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D012015:Reference Standards; D012988:Soil Microbiology
D004275:DNA, Ribosomal; D004247:DNA