Determination of the 3' terminal nucleotide of DNA fragments.
A new method for determining the 3'-terminal nucleotide of a DNA strand is presented. Use is made of the fact that one (and only one) 2',3'-dideoxyribonucleotide can be added to the 3'-end of a DNA fragment with calf thymus terminal transferase. Addition of more than one nucleotide analog per strand is impossible due to the absence of a 3'-terminal hydroxyl group. If the terminatind dideoxyribonucleotide contains an (alpha 32p) label, the resulting 3'-blocked strand can be digested by "nearest neighbor" techniques and the original 3'-endgroup determined. Picomole quantities of DNA strands can be labeled and the 3'-end determined.
D000818:Animals; D001483:Base Sequence; D001669:Biochemical Phenomena; D001671:Biochemistry; D002417:Cattle; D004247:DNA; D004254:DNA Nucleotidyltransferases; D003851:Deoxyribonucleases; D003854:Deoxyribonucleotides / Q000032:analysis; D008722:Methods; D008837:Micrococcus / Q000201:enzymology; D009713:Nucleotidyltransferases; D009841:Oligonucleotides; D010727:Phosphoric Diester Hydrolases; D011119:Polynucleotides; D013154:Spleen / Q000201:enzymology; D013950:Thymus Gland / Q000201:enzymology
D003854:Deoxyribonucleotides; D009841:Oligonucleotides; D011119:Polynucleotides; D004247:DNA; D004254:DNA Nucleotidyltransferases; D009713:Nucleotidyltransferases; D003851:Deoxyribonucleases; D010727:Phosphoric Diester Hydrolases