Line scanning system for direct digital chemiluminescence imaging of DNA sequencing blots

Journal Article · Thu Jul 01 00:00:00 EDT 1993 · Analytical Chemistry (Washington); (United States)

DOI:https://doi.org/10.1021/ac00061a024· OSTI ID:5695908

Karger, A E; Weiss, R; Gesteland, R F ^[1]

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1. Univ. of Utah, Salt Lake City (United States) Eccles Inst. of Human Genetics, Salt Lake City, UT (United States)

A cryogenically cooled charge-coupled device (CCD) camera equipped with an area CCD array is used in a line scanning system for low-light-level imaging of chemiluminescent DNA sequencing blots. Operating the CCD camera in time-delayed integration (TDI) mode results in continuous data acquisition independent of the length of the CCD array. Scanning is possible with a resolution of 1.4 line pairs/mm at the 50% level of the modulation transfer function. High-sensitivity, low-light-level scanning of chemiluminescent direct-transfer electrophoresis (DTE) DNA sequencing blots is shown. The detection of DNA fragments on the blot involves DNA-DNA hybridization with oligonucleotide-alkaline phosphatase conjugate and 1,2-dioxetane-based chemiluminescence. The width of the scan allows the recording of up to four sequencing reactions (16 lanes) on one scan. The scan speed of 52 cm/h used for the sequencing blots corresponds to a data acquisition rate of 384 pixels/s. The chemiluminescence detection limit on the scanned images is 3.9 [times] 10[sup [minus]18] mol of plasmid DNA. A conditional median filter is described to remove spikes caused by cosmic ray events from the CCD images. 39 refs., 9 refs.

• DOE Contract Number: FG02-88ER60700
• OSTI ID: 5695908
• Journal Information: Analytical Chemistry (Washington); (United States), Vol. 65:13; ISSN 0003-2700
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
37 INORGANIC
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
DNA
CHEMILUMINESCENCE
QUANTITATIVE CHEMICAL ANALYSIS
DIGITAL SYSTEMS
ENZYMES
IMAGE SCANNERS
CHEMICAL ANALYSIS
LUMINESCENCE
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
550200* - Biochemistry
550400 - Genetics
400100 - Analytical & Separations Chemistry