A novel method for preparing single-stranded DNA for pyrosequencing.
Pyrosequencing 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 are described. The method has also been fully automated.
Scottish Meningococcus and Pneumococcus Reference Laboratory, House on the Hill, Stobhill Hospital, Glasgow, G21 3UW, UK.
Matthew A
MA
Diggle
Stuart C
SC
Clarke
D000465:Algorithms; D004277:DNA, Single-Stranded / Q000235:genetics / Q000302:isolation & purification*; D020869:Gene Expression Profiling / Q000379:methods*; D006358:Hot Temperature; D009345:Neisseria meningitidis / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D018272:Porins / Q000235:genetics / Q000378:metabolism; D012371:Robotics / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*
D004277:DNA, Single-Stranded; D018272:Porins; C057699:porin protein, Neisseria