Octamer-primed sequencing technology: effects of dNTP supplementation.
Octamer-primed sequencing is a directed DNA sequencing strategy that employs the use of a presynthesized octamer primer library. Together with electronic octamer sequencing technology (eOST) it provides a faster, less expensive way to obtain DNA sequence information and can be used as a valuable tool for gap closure in large-scale genomic sequencing. In this paper we discuss the effect of dGTP/TTP supplementation on octamer sequencing. We show that addition of 75 microM dGTP and 5 microM TTP can improve the sequencing success rate by increasing the length and accuracy of generated sequence information. We also discuss the effect of template base composition immediately downstream of the octamer primer on the outcome of octamer sequencing.
Kraltcheva
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AI
Department of Biology and Biochemistry and Institute of Molecular Biology, University of Houston, Houston, TX 77204-5513, USA.
Hardin
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SH
D001482:Base Composition; D001483:Base Sequence; D004247:DNA / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics* / Q000378:metabolism; D003848:Deoxyguanine Nucleotides / Q000378:metabolism*; D015723:Gene Library; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009711:Nucleotides / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000191:economics / Q000379:methods*; D019914:Taq Polymerase / Q000378:metabolism; D013698:Templates, Genetic; D013942:Thymine Nucleotides / Q000378:metabolism*; D013997:Time Factors
D017931:DNA Primers; D003848:Deoxyguanine Nucleotides; D009711:Nucleotides; D009838:Oligodeoxyribonucleotides; D013942:Thymine Nucleotides; C029603:deoxyguanosine triphosphate; D004247:DNA; D019914:Taq Polymerase; C024157:thymidine 5'-triphosphate