Chain terminator sequencing of double-stranded DNA with built-in error correction.
Chain terminator sequencing is usually performed on single-stranded DNA. We propose a technique to determine a DNA sequence by sequencing both strands of a double strand in two four-lane gels. The first set of four lanes will contain a superposition of sequence information from the two complementary strands. The second set of four lanes will contain a different superposition in which one of the two single strands is "shifted" relative to the other. This technique provides redundant information sufficient for error correction--not just the error detection achieved by the standard approach of separately sequencing a single strand and its complement. The information content per four-lane gel is more than 50% higher in this double-stranded approach. Using this technique, the number of chain sequencings--including chain syntheses and gel separations--required to achieve a desired level of accuracy can be reduced by up to a factor of two. Unlike the ordinary single-stranded sequencing, some analysis is required to obtain the single-stranded sequence information from the double-stranded data, particularly in the presence of sequencing errors. This analysis and a computer simulation of double-stranded sequencing in the presence of errors are presented.
Miller
R L
RL
General Atomics, San Diego, CA 92186-9784.
Ohkawa
T
T
D000818:Animals; D003198:Computer Simulation; D004247:DNA / Q000737:chemistry*; D006801:Humans; D008433:Mathematics; D008957:Models, Genetic; D017422:Sequence Analysis, DNA / Q000379:methods*; D013728:Terminator Regions, Genetic / Q000235:genetics*
D004247:DNA