Novel separation and detection methods of DNA fragments in electrophoresis
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
- Resource Relation:
- Other Information: Thesis (Ph.D.)
- Country of Publication:
- United States
- Language:
- English
Similar Records
Novel separation and detection methods of DNA fragments in electrophoresis
Separation of DNA restriction fragments using capillary electrophoresis
Related Subjects
ORGANIC
PHYSICAL AND ANALYTICAL CHEMISTRY
59 BASIC BIOLOGICAL SCIENCES
DNA
DETECTION
ELECTROPHORESIS
CHARGE-COUPLED DEVICES
FLUORESCENCE
MOLECULAR BIOLOGY
SEPARATION PROCESSES
LUMINESCENCE
NUCLEIC ACIDS
ORGANIC COMPOUNDS
SEMICONDUCTOR DEVICES
400105* - Separation Procedures
550200 - Biochemistry