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DNA Res. 1995 Oct 31;2(5):225-7.

A non-radioactive DNA sequencing method using biotinylated dideoxynucleoside triphosphates and delta Tth DNA polymerase.

Author information

1
Tsuruga Institute of Biotechnology, Toyobo Co., Ltd., Fukui, Japan.

Abstract

We synthesized a set of four biotinylated dideoxynucleoside triphosphates (biotin-9-ddNTPs) and optimized the reaction conditions for non-radioactive cycle sequencing using modified Tth DNA polymerase (delta Tth) and a chemiluminescent detection system. The resulting sequencing ladders showed lower background compared to those with the conventional non-radioactive sequencing method which uses 5'-biotinylated primers, especially when PCR products were analysed. With our method, DNA sequences can be determined at any primer positions without preparing 5'-biotinylated primers for dideoxy chain-termination.

PMID:
8770564
DOI:
10.1093/dnares/2.5.225
[Indexed for MEDLINE]

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