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Sökning: WFRF:(Nyrén Pål) > (2015-2017)

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1.
  • Nordström, Tommy, et al. (författare)
  • Method enabling detection of single-stranded DNA ligation activity
  • 2017
  • Ingår i: Archives of industrial biotechnology. ; 1:1, s. 35-40
  • Tidskriftsartikel (refereegranskat)abstract
    • The enzyme DNA ligase is an essential tool in many different applications in the field of biotechnology. For these applications the DNA quality can be crucial for the final result. In this paper we present a new method that can be used for specific studies of DNA ligases and for simultaneous analyzes of DNA quality. More specifically, a simple, fast and sensitive method for detection of single-stranded DNA (ssDNA) ligase activity is presented. The new method is based on real-time detection of nucleotide incorporation over the ligation site catalyzed by a DNA polymerase. The ratio between the incorporation signal after and before the ligation site gives an estimate of the ligation efficiency. To further increase the robustness of the assay the exact amount of unligated product can be easily estimated in a separate assay using a 3’-end blocked oligonucleotide functioning as template. Produced circular DNA can be cleaned from unligated DNA by exonuclease I treatment and the quality of the circle can be easily analyzed by a simple sequencing procedure. If the ssDNA used for the ligation reaction contains truncated fragments this will be observed by uneven signals during the analysis. The new method can be a useful tool for researchers working with both basic and applied projects in the field of biotechnology.
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2.
  • Nyrén, Pål (författare)
  • The History of Pyrosequencing®
  • 2015
  • Ingår i: Methods in Molecular Biology. - New York, NY : Humana Press. - 1064-3745 .- 1940-6029. ; 1315, s. 3-15
  • Tidskriftsartikel (refereegranskat)abstract
    • One late afternoon in the beginning of January 1986, bicycling from the lab over the hill to the small village of Fulbourn, the idea for an alternative DNA sequencing technique came to my mind. The basic concept was to follow the activity of DNA polymerase during nucleotide incorporation into a DNA strand by analyzing the pyrophosphate released during the process. Today, the technique is used in multidisciplinary fields in academic, clinical, and industrial settings all over the word. This technique can be used for both single-base sequencing and whole-genome sequencing, depending on the format used.In this chapter, I give my personal account of the development of Pyrosequencing()-beginning on a winter day in 1986, when I first envisioned the method-until today, nearly 30 years later.
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Nyrén, Pål (1)
Nordström, Tommy (1)
Nyrén, Pål, 1955- (1)
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