Opening book details…
Can I read Method for preparing single-stranded DNA templates for Pyrosequencing using vector ligation and universal biotinylated primers on EtoBox?
Method for preparing single-stranded DNA templates for Pyrosequencing using vector ligation and universal biotinylated primers by Marco Groth; Klaus Huse; Kathrin Reichwald; Stefan Taudien; Jochen Hampe; Philip Rosenstiel; Gerd Birkenmeier; Stefan Schreiber; Matthias Platzer is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is Method for preparing single-stranded DNA templates for Pyrosequencing using vector ligation and universal biotinylated primers about?
In Pyrosequencing, the addition of nucleotides to a primer-template hybrid is monitored by enzymatic conversion of chemical energy into detectable light. The technique yields both qualitative and quantitative sequence information because the chemical energy is released by a stoichiometric split off of pyrophosphates from incorporated deoxynucleotide triphosphates and a defined nucleotide dispensation order is given. Because Pyrosequencing works best if single-stranded DNA templates are used, template generation usually requires PCR with a target-specific biotinylated primer and a subsequent purification involving interaction of the biotin label with immobilized streptavidin. To circumvent the need for numerous and expensive template-specific biotinylated primers, we developed a method that uses the ligation of amplified DNA fragments into a plasmid vector, thereby facilitating subsequent PCR using a universal vector-specific biotinylated primer. This approach allows easy and straightforward isolation of single-stranded templates of any PCR product. As a proof of principle, we used the method for genotyping two single-nucleotide polymorphisms in the human genes CARD15 and A2M and fo
Who reads Method for preparing single-stranded DNA templates for Pyrosequencing using vector ligation and universal biotinylated primers?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Marco Groth; Klaus Huse; Kathrin Reichwald; Stefan Taudien; Jochen Hampe; Philip Rosenstiel; Gerd Birkenmeier; Stefan Schreiber; Matthias Platzer
- Publisher
- Elsevier Science; Elsevier ; Elsevier Inc.; Elsevier BV (ISSN 0003-2697)
- Published
- 2006
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)