branded 3' UHEULHET One dNTP is a dctP dTTP 4 reaction ekEs ::::: HA'TGCGAGACT dGCGAGACT T Gelelectrophoresis ddGATGCGAGACT ddGACACT dC GATGCGAGACI Reading sequence bMom to topi-A-G-C-G-T-A-G-C Its complement b the ortal teple Maid (-sB-T-C-G-C-A-T-C-G
) 5 OH ② PI( OH OH Primer strand A T G CC G 3′ AC G TA C T Template strand O-P-0-P-0-P-0-cH2O Base O HH ddNTP Analog b
Colored Primer dye label Template of unknown sequence DNA polymerase 4 dNTPs, one ddNTP (C) C C C C Denature C C Dye-labeled segments of DNA C spied from CoI C template with C unknown sequen
ddATP ddttP ddGtP ddcTP DNA DNA DNA DNA dNTPs dNTPs dNTPs dNTPs 5 TA TAGTA Each fragment is terminated by a dda TAGTACA TAGTACAGTA TAGTACAGTAGTTCA TAGTACAGTAGTTCAGA
5’ 3’ ||||||||||||||| ddATP DNA dNTPs ddTTP DNA dNTPs ddGTP DNA dNTPs ddCTP DNA dNTPs A T C A T G T C A T C A A G T C T A G C A C T A T A G T A T A G T A C A T A G T A C A G T A T A G T A C A G T A G T T C A T A G T A C A G T A G T T C A G A • Each fragment is terminated by a ddA
Primer 5 OH 3 CTAAGOTCGACT Template dcTP. dGTP dATP dTTP + ddATP ddCTP + ddGTP t ddTTP GATTCGAGCTGddA GATTCGAGddC GATTCGAGCTddG GATTCGAGCddT GATTCGddA GATTddc GATrCGAddG GATed GodA GATTCddG GAdd ddG C G T 11 10 987654321 AGTcGAGcTTA