Sequencing BAC DNA With Near-Infrared
Fluorescent Non-Nucleotide Terminators

Stephen C. Roemer, Vince R. Boveia, Craig M. Johnson and D. Michael Olive
LI-COR Inc., Biotechnology, 4647 Superior Street, Lincoln, NE 68504

Sequencing Results

Comparison of the performance of IRD800-labeled acyNTPs and IRD800-labeled ddNTP for DNA sequencing. The lane-to-lane band intensities for all four IRD800-labeled acyNTPs were equivalent to those obtained with IRD800-labeled ddNTPs throughout the sequence ladder.

Comparison of read length and accuracy to 1000 bases for two primer walking projects
using either dye-primer or dye-AcycloTerminator sequencing.

	Dye-AcycloTerminator		Dye-Primer
Custom Primer	Read Length	Accuracy (%)	Read Length	Accuracy (%)
Template p4CG9
T3	1157	99.1	1067	99.4
T7	1090	99.4	1099	99.8
967T7F	1126	98.9	1163	100.0
218T7R-1	1192	98.4	1120	99.9
914T3F	1059	99.4	1129	99.8
284T3R	1093	99.4	1120	100.0
1872T7F	1090	99.8	1174	100.0
1874T3F	1162	99.5	1063	99.8
310T7R	1020	98.4	1154	99.6
221T3R	1120	99.5	1138	99.9
2742T3F	1118	97.6	1039	99.9
251T3R	1178	99.2	1140	99.9
2844T7F	1083	98.8	1085	99.8
218T7R-2	1010	98.2	1083	99.6
3624T3F	1177	99.4	1147	99.4
211T3R	1066	99.6	1224	99.9
3701T7F	1121	99.4	1112	99.6
228T7R	1036	98.0	1053	99.4
Template pUC19
M13F	1005	98.9	1092	99.5
508F	1073	99.9	1198	99.8
928F	1062	98.9	1164	99.7
1243F	1093	99.1	1108	100.0
1585F	1040	99.2	1184	99.6
1991F	1040	99.4	1141	99.5
2286F	1037	99.4	1152	99.6
2482F	1116	99.7	1067	98.1
MEAN	1090	99.1	1123	99.7