Specific Binding of a Synthetic Oligonucleotide to the Yeast Iso-1 Cytochrome C̲ MRNA and Gene |
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5'-end labeled acetic acid added addition agarose gel amount assay bands base binding carrying cDNA cells cellulose acetate Chemical chloride chloroform chromatography column complementary complete concentration containing cytochrome c mRNA cytoplasm DEAE-cellulose deletion determined developed digest dissolved dried Eco RI eluted enzyme et al ethanol ether evaporated excess experiments filter followed Fractions fragments fully protected gene hybridization incubated initiation codon iso-1 cytochrome isolated labeled labeled 15-mer LEGEND FOR FIGURE loaded mapping marker material method mmole molecule mRNA hybrid mutants nucleotides oligonucleotide origin partial phosphate plate polymerase precipitated prepared presence primer extension probe procedure protein purchased purification pyridine reaction mixture reagents region remove ribosome binding rRNA selective separated Sephadex Sequence Analysis showed shown silica gel solution specific strain structure synthesis TABLE tion trinucleotide washed wild type yeast DNA yield