RT Journal
A1 Schmalzing, Dieter
A1 Tsao, Norman
A1 Koutny, Lance
A1 Chisholm, Dan
A1 Srivastava, Alok
A1 Adourian, Aram
A1 Linton, Lauren
A1 McEwan, Paul
A1 Matsudaira, Paul
A1 Ehrlich, Daniel
T1 Toward Real-World Sequencing by Microdevice Electrophoresis
JF Genome Research 
JO Genome Research 
YR 1999 
FD September 01 
VO 9 
IS 9 
SP 853 
OP 858 
DO 10.1101/gr.9.9.853 
UL http://genome.cshlp.org/content/9/9/853.abstract 
AB We report results using a microdevice for DNA sequencing using samples from chromosome 17, obtained from the Whitehead Institute Center for Genome Research (WICGR) production line. The device had an effective separation distance of 11.5 cm and a lithographically defined injection width of 150 μm. The four-color raw data were processed, base-called by the sequencing software Trout, and compared to the corresponding ABI 377 sequence from WICGR. With a criteria of 99% accuracy, we achieved average continuous reads of 505 bases in 27 min with 3% linear polyacrylamide (LPA) at 150 V/cm, and 460 bases in 22 min with 4% LPA at 200 V/cm at a temperature of 45°C. In the best case, up to 565 bases could be base-called with the same accuracy in <25 min. In some instances, Trout allowed for accurate base-calling down to a resolution R as low as R = 0.35. This may be due in part to the high signal-to-noise ratio of the microdevice. Unlike many results reported on capillary machines, no additional sample cleanup other than ethanol precipitation was required. In addition, DNA fragment biasing (i.e., discrimination against larger fragments) was reduced significantly through the unique sample injection mechanism of the microfabricated device. This led to increased signal strength for long fragments, which is of great importance for the high performance of the microdevice.