Novel separation and detection methods of DNA fragments in electrophoresis

Thesis/Dissertation · Wed Jan 01 04:00:00 EST 1992

OSTI ID:7102328

Chan, K C

A charge-coupled device (CCD) based electrophoresis system was developed. The system allowed non-destructive, sensitive, and on-line detection of native DNA in slab-gel electrophoresis via ultraviolet absorption measurement. The detection limit of double-stranded DNA fragment was 5 ng per band. Since the amount of DNA used in this experiment was typical, the CCD-based system could be readily implemented in molecular biology. Gel-filled and non-gel sieving capillary electrophoresis (CE) was developed for rapid and efficient separation of double-stranded DNA fragments. For the gel-filled CE separation a new gel matrix, the HydroLink gel (HL), was used. The HL capillary gel was easier to cast than the polyacrylamide capillary gel. For the non-gel separation, a GC capillary was used as the separation chamber, and cellulose additive was included in the electrophoresis as the sieving medium. Indirect fluorometry was applied in non-gel and gel electrophoresis for the detection of DNA fragments. This method allowed nondestructive and on-line detection of DNA during electrophoresis. The amount of DNA used with this method was comparable to those obtained with absorption measurement.

• Research Organization: Iowa State Univ. of Science and Technology, Ames, IA (United States)
• OSTI ID: 7102328
• Country of Publication: United States
• Language: English

Related Subjects

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
400105* -- Separation Procedures
550200 -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
CHARGE-COUPLED DEVICES
DETECTION
DNA
ELECTROPHORESIS
FLUORESCENCE
LUMINESCENCE
MOLECULAR BIOLOGY
NUCLEIC ACIDS
ORGANIC COMPOUNDS
SEMICONDUCTOR DEVICES
SEPARATION PROCESSES