Quantitation of mixed-base populations of HIV-1 variants by automated DNA sequencing with BODIPY dye-labeled primers
Direct DNA sequencing of human immunodeficiency virus type 1 (HIV-1) pol and env gene regions was characterized for accuracy and precision. Restricted maximum likelihood (REML) analysis of molecular clone reconstruction experiments using a primer-walking strategy showed that the BODIPY and BODIPY en...
| Veröffentlicht in: | BioTechniques. - 1993. - 25(1998), 3 vom: 07. Sept., Seite 446-7, 450-2, 454, passim |
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| 1. Verfasser: | |
| Weitere Verfasser: | , , , |
| Format: | Aufsatz |
| Sprache: | English |
| Veröffentlicht: |
1998
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| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Journal Article 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene Boron Compounds DNA Primers DNA, Viral HIV Envelope Protein gp120 DNA-Directed DNA Polymerase EC 2.7.7.7 |
| Zusammenfassung: | Direct DNA sequencing of human immunodeficiency virus type 1 (HIV-1) pol and env gene regions was characterized for accuracy and precision. Restricted maximum likelihood (REML) analysis of molecular clone reconstruction experiments using a primer-walking strategy showed that the BODIPY and BODIPY energy-transfer (BET) dye primers sets were significantly more accurate in quantitating heterogenous base mixtures than the fluorescein/rhodamine dye primers. Of the three sets examined, the BET dye primers were the most accurate. The improved accuracy correlated with the reduced emission band-widths of BODIPY and BET dye primers and the more uniform signal intensities of BET dye primers. However, comparing % coefficients of variation (CV) for the three dye primer sets, revealed that BODIPY dye primers gave better precision than both BET and fluorescein/rhodamine dye primer sets. Several sequence-dependent motifs were identified that showed specific nucleotide-biased incorporation and were determined to be the major variable component of the total %CV. Taken together, these results show that BODIPY and BET direct DNA sequencing can accurately and precisely characterize complex mixed-base populations |
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| Beschreibung: | Date Completed 25.11.1998 Date Revised 21.11.2008 published: Print Citation Status MEDLINE |
| ISSN: | 0736-6205 |