Nucleic Acids Research, 1981, Vol. 9, No. 4 879-894
© 1981
MOLECULAR BIOLOGY |
The use of synthetic oligonucleotides as hybridization probes. II. Hybridization of oligonucleotides of mixed sequence to rabbit ß-globin DNA
Division of Biology, Molecular Genetics Section, City of Hope Research Institute Duarte, CA 91010, USA *Pharmaceutical Institute, School of Medicine, Keio University Tokyo, Japan
Received December 1, 1980. Two oligonucleotides 14-bases long were synthesized, one complementary to rabbit ß-globin DNA (RßG14A) and the other with the same sequence except for a single base change (T for C) (RßG14B) Hybridization conditions were established such that RRßG14A would hybridize to globin DNA while RßG14B would not. We also synthesized a mixture of 13-base long oligonucleotides (RßG13Mix), representing eight of the possible coding sequences for amino acids 15–19 of rabbit ß-globin. One of the eight is complementary to globin DNA. RßG13Mix was found to hybridize specifically to globin DNA under conditions where oligonucleotides forming single base pair mismatches do not. Furthermore, RßGl3Mix was shown to hybridize specifically to colonies containing a plasmid with a globin DNA insert. These results are discussed with respect to a general procedure for screening recombinant clones for those containing DNA coding for a protein of known amino acid sequence.
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