Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1989-02-15
1994-10-04
Fleisher, Mindy B.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
536 253, 935 77, 935 78, C12Q 168, C07H 2100, C12N 1500
Patent
active
053525787
ABSTRACT:
A method of purifying full length synthetic target oligonucleotides from a mixture of oligonucleotides, particularly from a mixture containing truncated or failed sequences is disclosed. The method involves attaching a short nucleotide sequence complementary to the 5' end of a target oligonucleotide to a solid support. The complementarity between the most 5' nucleotides of the target oligonucleotide and the bound oligonucleotide results in hybridization which serves to retain the target oligonucleotide. Truncated or failed sequences lacking 5' sequences complementary to the attached oligonucleotide, fail to hybridize and therefore are not retained. The method makes it possible to purify gram quantities of synthetic deoxyribonucleic acids or ribonucleic acids and sequences which have modifications, such as on the phosphate backbone. The support-bound nucleotide sequences are stable under conditions of purification and therefore can be reused.
REFERENCES:
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Aviv and Leder, "Purification of Biologically Active Globin Messenger RNA by Chromatography on Oligothymidylic Acid-Cellulose", Proc. Natl. Acad. Sci. USA, 69: 1408-1412 (1972).
Okamura et al., "Purification of Terminal Deoxynucleotidyl-transferase by Olignoncleotide Affinity Chromatography", J. Bio. Chem. , 253: 3765-3767 (1978).
Thanki et al., "Selective Binding of Mouse Estradiol-Roceptor Complexes to Oligo(dT)-Cellulose," J. Biol. Chem. 253: 7744-7750 (1978).
Sheldon et al., "Detection o Polyadenylic Acid Sequences in Viral and Eukaryotic RNA", Proc. Nat. Acad. Sci. USA, 69: 417-421 (1972).
Agrawal Sudhir
Zamecnik Paul C.
Fleisher Mindy B.
Worcester Foundation for Experimental Biology
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