8 research outputs found
Reading Single DNA with DNA Polymerase Followed by Atomic Force Microscopy
The importance of DNA sequencing in the life sciences and personalized medicine is continually increasing. Single-molecule sequencing methods have been developed to analyze DNA directly without the need for amplification. Here, we present a new approach to sequencing single DNA molecules using atomic force microscopy (AFM). In our approach, four surface conjugated nucleotides were examined sequentially with a DNA polymerase immobilized AFM tip. By observing the specific rupture events upon examination of a matching nucleotide, we could determine the template base bound in the polymerase's active site. The subsequent incorporation of the complementary base in solution enabled the next base to be read. Additionally, we observed that the DNA polymerase could incorporate the surface-conjugated dGTP when the applied force was controlled by employing the force-clamp mode.X1114Ysciescopu
Telbivudine versus entecavir in patients with undetectable hepatitis B virus DNA: a randomized trial
Reading Single DNA with DNA Polymerase Followed by Atomic Force Microscopy
The
importance of DNA sequencing in the life sciences and personalized
medicine is continually increasing. Single-molecule sequencing methods
have been developed to analyze DNA directly without the need for amplification.
Here, we present a new approach to sequencing single DNA molecules
using atomic force microscopy (AFM). In our approach, four surface-conjugated
nucleotides were examined sequentially with a DNA polymerase-immobilized
AFM tip. By observing the specific rupture events upon examination
of a matching nucleotide, we could determine the template base bound
in the polymerase’s active site. The subsequent incorporation
of the complementary base in solution enabled the next base to be
read. Additionally, we observed that the DNA polymerase could incorporate
the surface-conjugated dGTP when the applied force was controlled
by employing the force-clamp mode