DNA sequencing using fluorescence background electroblotting membrane

Number of patents in Portfolio can not be more than 2000

United States of America Patent

PATENT NO 5112736
SERIAL NO

07365693

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ATTORNEY / AGENT: (SPONSORED)

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Abstract

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A method for the multiplex sequencing on DNA is disclosed which comprises the electroblotting or specific base terminated DNA fragments, which have been resolved by gel electrophoresis, onto the surface of a neutral non-aromatic polymeric microporous membrane exhibiting low background fluorescence which has been surface modified to contain amino groups. Polypropylene membranes are preferably and the introduction of amino groups is accomplished by subjecting the membrane to radio or microwave frequency plasma discharge in the presence of an aminating agent, preferably ammonia. The membrane, containing physically adsorbed DNA fragments on its surface after the electroblotting, is then treated with crosslinking means such as UV radiation or a glutaraldehyde spray to chemically bind the DNA fragments to the membrane through said smino groups contained on the surface thereof. The DNA fragments chemically bound to the membrane are subjected to hybridization probing with a tagged probe specific to the sequence of the DNA fragments. The tagging may be by either fluorophores or radioisotopes. The tagged probes hybridized to said target DNA fragments are detected and read by laser induced fluorescence detection or autoradiograms. The use of aminated low fluorescent background membranes allows the use of fluorescent detection and reading even when the available amount of DNA to be sequenced is small. The DNA bound to the membrances may be reprobed numerous times.

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Patent Owner(s)

Patent OwnerAddressTotal Patents
BRIGHAM YOUNG UNIVERSITYPROVO, UT233
UNIVERSITY OF UTAHSALT LAKE CITY, UT905

International Classification(s)

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Inventor(s)

Inventor Name Address # of filed Patents Total Citations
Caldwell, Karin D Salt Lake City, UT 15 881
Chu, Tun-Jen Salt Lake City, UT 12 611
Pitt, William G Orem, UT 10 229

Cited Art Landscape

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