Fluorescence-detected DNA sequencing

Technical Report · Mon Jan 01 04:00:00 EST 1990

DOI:https://doi.org/10.2172/5619036· OSTI ID:5619036

Haugland, R P

Our research effort funded by this grant primarily focused on development of suitable fluorescent dyes for DNA sequencing studies. Prior to our efforts, the dyes being sued in commercial DNA sequencers were various versions of fluorescein dyes for the shorter wavelengths and of rhodamine dyes for the longer wavelengths. Our initial goal was to synthesize a set of four dyes that could all be excited by the 488 and 514 nm line of the argon laser lines and that have emission spectra that minimize spectral overlap. The specific result sought was higher fluorescent intensity, particularly of the longest wavelength dyes than was available using existing dyes. Another important property of the desired set of dyes was uniform ionic charge in order to have minimum interference on the electrophoretic mobility during the sequencing. During the period of this grant we prepared and characterized four types of dyes: fluorescent bifluorophores, derivatives of rhodamine dyes, derivatives of rhodol dyes and derivatives of boron dipyrromethene difluoride (BODIPY{trademark}) dyes.

• Research Organization: Molecular Probes, Inc., Eugene, OR (United States)
• Sponsoring Organization: DOE; USDOE, Washington, DC (United States)
• DOE Contract Number: FG06-88ER60684
• OSTI ID: 5619036
• Report Number(s): DOE/ER/60684-T1; ON: DE92011124
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AROMATICS
CARBOXYLIC ACIDS
DNA SEQUENCING
DOCUMENT TYPES
DYES
FLUORESCEIN
FLUORESCENCE
HYDROXY ACIDS
HYDROXY COMPOUNDS
LUMINESCENCE
ORGANIC ACIDS
ORGANIC COMPOUNDS
PHENOLS
POLYPHENOLS
PROGRESS REPORT
STRUCTURAL CHEMICAL ANALYSIS
SYNTHESIS