| Autor: |
Walker, John M., Goulden, Nicholas J., Steward, Colin G., Dunn, Paul P. J., Day, Sarinder, Williams, Siaron, Bendukidze, Nina |
| Zdroj: |
Pediatric Hematology; 2004, p233-246, 14p |
| Abstrakt: |
The ever increasing numbers of reported HLA alleles pose a problem to the modern tissue typing laboratory (Table 1) (1-4). PCR-sequencing based typing (PCR-SBT) analyzes the complete nucleotide sequence of the polymorphic exons of HLA class I and II and, as such, provides the highest resolution available for HLA typing which includes the complete characterization of new alleles. Sequencing using Taq DNA polymerase, or "cycle sequencing," is essentially PCR in which one primer and the enzyme have been modified to ensure even incorporation of each ddNTP into the growing DNA strand. Such even incorporation is important for the detection of heterozygous bases where two different nucleotides are present. The methods described here have been developed and used on a slab gel automated DNA sequencer (Applied Biosystems 373) and they are now being used on a 16-capillary automated DNA sequencer (Applied Biosystems 3100 Genetic Analyzer). [ABSTRACT FROM AUTHOR] |
| Databáze: |
Supplemental Index |
| Externí odkaz: |
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