Novel separation and detection methods of DNA fragments in electrophoresis

Technical Report · Wed Jan 27 04:00:00 EST 1993

OSTI ID:7063614

Chan, King Cheung

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 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 non-destructive 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: Ames Lab., IA (United States)
• Sponsoring Organization: DOE; USDOE, Washington, DC (United States)
• DOE Contract Number: W-7405-ENG-82
• OSTI ID: 7063614
• Report Number(s): IS-T-1596; ON: DE93007539
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
CHARGE-COUPLED DEVICES
CHROMATOGRAPHY
DNA
ELECTROPHORESIS
EMISSION SPECTROSCOPY
FLUORESCENCE SPECTROSCOPY
GAS CHROMATOGRAPHY
NUCLEIC ACIDS
OLIGONUCLEOTIDES
ORGANIC COMPOUNDS
SEMICONDUCTOR DEVICES
SEPARATION PROCESSES
SPECTROSCOPY