Use of non-cross-linked polyacrylamide for four-color DNA sequencing by capillary electrophoresis separation of fragments up to 640 bases in length in two hours

Journal Article · Fri Dec 15 00:00:00 EST 1995 · Analytical Chemistry (Washington)

DOI:https://doi.org/10.1021/ac00120a026· OSTI ID:171374

Zhang, J Z; Fang, Y; Hou, J Y; Ren, H J; Jiang, R; Roos, P; Dovichi, N J ^[1]

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1. Univ. of Alberta, Edmonton (Canada)

Four-color DNA cycle sequencing was performed on an M13mp18 template using dye-labeled primers. Sequencing fragments were separated by capillary electrophoresis at 60{degree}C and at an electric field of 150 V/cm. The sieving medium was 5%T, non-cross-linked polyacrylamide in 7M urea. The use of high temperature for the separation reduces formation of secondary structures in the sequencing fragments, generating a sequence that is free of compressions without the use of strongly denaturing gels. The use of high temperatures also increases the separation rate compared with room-temperature operation. Fragments up to 640 bases are separated in less than 2 h. 27 refs., 4 figs.

• Sponsoring Organization: USDOE
• DOE Contract Number: FG02-91ER61123
• OSTI ID: 171374
• Journal Information: Analytical Chemistry (Washington), Vol. 67, Issue 24; Other Information: PBD: 15 Dec 1995
• Country of Publication: United States
• Language: English

Related Subjects

36 MATERIALS SCIENCE
40 CHEMISTRY
55 BIOLOGY AND MEDICINE
BASIC STUDIES
DNA
DNA SEQUENCING
CHEMICAL ANALYSIS
POLYMERS
POLYAMIDES
CAPILLARY FLOW
ELECTROPHORESIS
TEMPERATURE DEPENDENCE