Process for producing cDNA

Details Process for producing cDNA Process for producing cDNA

1994-07-25

1998-06-02

Jones, W. Gary

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 6, 435 5, 435 912, 4351739, 42218601, 42218603, 204156, 204554, 436125, 436126, 536 243, 536 2432, 536 2433, C12P 1934, C12Q 168, C12Q 170, B01J 1908

Patent

active

057598207

ABSTRACT:
The process includes the steps of: a) contacting a liquid containing mRNA with an insoluble support having DNA probes attached thereto via the 5' -terminus thereof whereby the MRNA is hybridized to said probes and hence to said support; b) removing said liquid; and c) adding enzymes and nucleotides in solution whereby the probe functions as a primer to produce single stranded cDNA on the mRNA probe. The probe DNA may be oligo-dT or a specific DNA sequence, such as one that is complementary to a conserved region in a class of mRNA molecules. The insoluble support comprises magnetic particles which are monodisperse polymer particles comprising superparamagnetic iron oxide, a coating to reduce non-specific binding and a substituent for attaching an oligonucleotide.

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Kuribayashi et al., "A rapid and efficient purification of poly(A)-mRNA by oligo(dT).sub.30 -Latex", Nucleic Acids Research, Symposium Series No. 18, pp. 61-64 (1988).
Atkinson et al., "A convenient procedure for the synthesis of oligodeoxyribonucleotide affinity columns for the isolation of mRNA", Nucleic Acids Research, 16:6232 (1988).
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