Reliable and efficient direct sequencing of PCR-amplified double-stranded genomic DNA template
| Autor: | Mira Jung, A Dritschilo, U Kasid |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
DNA nanoball sequencing
Base Sequence Molecular Sequence Data Sequence assembly DNA DNA Neoplasm Computational biology Ion semiconductor sequencing Biology Polymerase Chain Reaction Introns Sequencing by ligation chemistry.chemical_compound Subcloning Sequencing by hybridization chemistry Evaluation Studies as Topic Dideoxynucleotide Genetics Humans Genomic library DNA Probes Genetics (clinical) |
| Zdroj: | Genome Research. 1:171-174 |
| ISSN: | 1088-9051 |
| DOI: | 10.1101/gr.1.3.171 |
| Popis: | Modified PCR amplification and direct sequencing procedures for the double-stranded genomic DNA template are described. Advantages of the approach we describe are: background artifact bands previously observed using high-molecular-weight DNA as a template were eliminated by this protocol; no gel purification or subcloning of the PCR-amplified double-stranded fragment was required prior to direct sequencing; and sequences of 300 nucleotides can be easily read even after a single loading. The successful use of the modified dideoxynucleotide chain-termination method for direct sequencing of both strands demonstrates the efficiency of this technique for removing sequencing artifacts and for producing reliable sequence data. |
| Databáze: | OpenAIRE |
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