A simple DNA stretching method for fluorescence imaging of single DNA molecules.
Stretching or aligning DNA molecules onto a surface by means of molecular combing techniques is one of the critical steps in single DNA molecule analysis. However, many of the current studies have focused on lambda-DNA, or other large DNA molecules. There are very few studies on stretching methodologies for DNA molecules generated via PCR (typically smaller than 20 kb). Here we describe a simple method of stretching DNA molecules up to 18 kb in size on a modified glass surface. The very low background fluorescence allows efficient detection of single fluorescent dye labels incorporated into the stretched DNA molecules.
Chan
Ting-Fung
TF
Cardiovascular Research Institute, University of California, San Francisco CA, USA.
Ha
Connie
C
Phong
Angie
A
Cai
Dongmei
D
Wan
Eunice
E
Leung
Lucinda
L
Kwok
Pui-Yan
PY
Xiao
Ming
M
D004247:DNA / Q000032:analysis* / Q000737:chemistry; D005456:Fluorescent Dyes; D005898:Glass / Q000737:chemistry; D008856:Microscopy, Fluorescence; D016133:Polymerase Chain Reaction; D013194:Staining and Labeling / Q000379:methods
D005456:Fluorescent Dyes; D004247:DNA