A novel method for preparing single-stranded DNA for Pyrosequencing™

被引：12

作者：

Diggle, MA

Clarke, SC

机构：

[1] Scottish Meningococcus & Pneumococcus Reference L, Glasgow G21 3UW, Lanark, Scotland

[2] Univ Glasgow, Fac Biomed & Life Sci, Glasgow G12 8QQ, Lanark, Scotland

来源：

MOLECULAR BIOTECHNOLOGY | 2003年 / 24卷 / 02期

关键词：

automation; genotyping; nucleotide sequencing; polymerase chain reaction; Pyrosequencing (TM); single-stranded DNA;

D O I：

10.1385/MB:24:2:221

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Pyrosequencing(TM) technology is a powerful genotyping tool that requires the generation of single-stranded DNA. Currently, two simple, solid-phase-based methods are available for this, but they require special equipment, they are not automated, and they are relatively expensive because of the need for biotinylated polymerase chain reaction primers. In this article, an enzymatic liquid-phase method for the generation of high-quality, single-stranded DNA, and its novel use for Pyrosequencing(TM) are described. The method has also been fully automated.

引用

收藏

页码：221 / 224

页数：4

相关论文

共 10 条

[1] Molecular techniques: divide or share [J].

Boxer, M .

JOURNAL OF CLINICAL PATHOLOGY, 2000, 53 (01) :19-21

[2] Nucleotide sequence-based typing of bacteria and the impact of automation [J].

Clarke, SC .

BIOESSAYS, 2002, 24 (09) :858-862

[3] Automated PCR/sequence template purification [J].

Clarke, SC ;

Diggle, MA .

MOLECULAR BIOTECHNOLOGY, 2002, 21 (03) :221-224

[4] Semiautomation of multilocus sequence typing for the characterization of clinical isolates of Neisseria meningitidis [J].

Clarke, SC ;

Diggle, MA ;

Edwards, GFS .

JOURNAL OF CLINICAL MICROBIOLOGY, 2001, 39 (09) :3066-3071

[5] What a load of old sequence!!! [J].

Diggle, MA ;

Clarke, SC .

JOURNAL OF CLINICAL MICROBIOLOGY, 2002, 40 (07) :2707-2707

[6]

Diggle MA, 2001, REV MED MICROBIOL, V12, P211

[7] Rapid molecular identification and subtyping of Helicobacter pylori by pyrosequencing of the 16S rDNA variable V1 and V3 regions [J].

Monstein, HJ ;

Nikpour-Badr, S ;

Jonasson, J .

FEMS MICROBIOLOGY LETTERS, 2001, 199 (01) :103-107

[8] Principles and applications of methods for DNA-beased typing of microbial organisms [J].

Olive, DM ;

Bean, P .

JOURNAL OF CLINICAL MICROBIOLOGY, 1999, 37 (06) :1661-1669

[9] Pyrosequencing sheds light on DNA sequencing [J].

Ronaghi, M .

GENOME RESEARCH, 2001, 11 (01) :3-11

[10] Pyrosequencing as a method for grouping of Listeria monocytogenes strains on the basis of single-nucleotide polymorphisms in the inlB gene [J].

Unnerstad, H ;

Ericsson, H ;

Alderborn, A ;

Tham, W ;

Danielsson-Tham, ML ;

Mattsson, JG .

APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 2001, 67 (11) :5339-5342