Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1997-01-16
1998-12-22
Jones, W. Gary
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
536 221, 536 242, 536 2453, 536 2534, 536 254, 435 9152, 435395, 435810, 435 71, 435 9151, 424 858, C07H 1900, C07H 2104, C07H 2100, C12P 1934
Patent
active
058518053
ABSTRACT:
An improved method for amplifying cDNA fragments corresponding to the mRNA species expressed in cells is described. The method involves priming the mRNA with a first labeled synthetic oligodeoxynucleotide and then reverse transcribing the mRNA to produce a mRNA:cDNA hybrid; and then producing a double-stranded DNA from the hybrid. This DNA is digested with a restriction endonuclease and then a synthetic DNA linker, carrying a second label, is ligated at the cut to the DNA strand containing the 5' phosphate group. The DNA is then isolated with a first reagent which binds the second label. The DNA fragments of interest are then isolated using a second reagent which binds the first label. The isolated DNA fragments are amplified using polymerase chain reaction, and then separated on a sequencing gel. The method is particularly useful for comparing the mRNA species expressed in the cells of various samples.
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Maher Veronica M.
McCormick Joseph Justin
Wei Dong
Board of Trustees operating Michigan State University
Jones W. Gary
McLeod Ian C.
Tung Joyce
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