Chain Terminator Sequencing of Double-stranded DNA with Built-in Error Correction
✍ Scribed by R.L. Miller; T. Ohkawa
- Publisher
- Elsevier Science
- Year
- 1993
- Tongue
- English
- Weight
- 726 KB
- Volume
- 161
- Category
- Article
- ISSN
- 0022-5193
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✦ Synopsis
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-noi 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.