Detection and isolation of glucagon mRNA using a synthetic oligo

Details Detection and isolation of glucagon mRNA using a synthetic oligo Detection and isolation of glucagon mRNA using a synthetic oligo

1982-04-08

1983-08-30

Wiseman, Thomas G.

Chemistry: molecular biology and microbiology

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

Preparing compound containing saccharide radical

435 6, 435 68, 435172, 536 29, 436 94, C12Q 168, C12P 1934, C12P 2100, C12N 1500

Patent

active

044017596

ABSTRACT:
A synthetic oligodeoxynucleotide complementary to glucagon mRNA and a method of using it to detect and isolate glucagon mRNA and cDNA from human and rabbit pancreas. A unique 14 base oligodeoxynucleotide dTTCATCAGCCACTG wherein T represents thymine, G represents guanine, A represents adenine, and C represents cytosine and where at least 13 of the 14 nucleotides are as indicated (one of those indicated may be replaced by one of the other three mentioned nucleotides) has been found to be complementary to glucagon mRNA. To isolate glucagon mRNA, total RNA is first extracted from human or rabbit pancreas and A+ RNA (mRNA-poly A) is isolated from the total RNA. The A+ RNA is then treated with the oligodeoxynucleotide, and the resulting hybridized RNA is enzymatically converted to glucagon mRNA which can then be purified, copied into glucagon cDNA and used in a conventional manner to produce glucagon by cloning technique.

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Ryndich, et al., "Electron Microscopic Study of the DNA in Products of Reverse Transcription", Chem. Absts. vol. 91, No. 1, p. 160 (1979), Abs. No. 1500d.

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