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ZB 10 - Soft Condensed Matter (R. Holyst)

We are different, but we all do great Science; and we have a lot of fun doing it!
We are different, but we all do great Science; and we have a lot of fun doing it!
We are different, but we all do great Science; and we have a lot of fun doing it!
We are different, but we all do great Science; and we have a lot of fun doing it!
We are different, but we all do great Science; and we have a lot of fun doing it!
We are different, but we all do great Science; and we have a lot of fun doing it!
We are different, but we all do great Science; and we have a lot of fun doing it!
We are different, but we all do great Science; and we have a lot of fun doing it!

Publication

A flexible fluorescence correlation spectroscopy based method for of the DNA double labeling efficiency with precision control

Author(s): Hou, Sen and Tabaka, Marcin and Sun, Lili and Trochimczyk, Piotr and Tomasz S. and Kalwarczyk, Tomasz and Zhang, Xuzhu and Holyst, Robert
Title: A flexible fluorescence correlation spectroscopy based method for of the DNA double labeling efficiency with precision control
Abstract: We developed a laser-based method to quantify the double labeling of double-stranded DNA (dsDNA) in a fluorescent dsDNA pool fluorescence correlation spectroscopy (FCS). Though, for biochemistry, accurate measurement of this parameter is of importance, before our work it was almost impossible to what percentage of DNA is doubly labeled with the same dye. The is produced by annealing complementary single-stranded DNA (ssDNA) with the same dye at 5' end. Due to imperfect ssDNA labeling, resulting dsDNA is a mixture of doubly labeled dsDNA, singly labeled and unlabeled dsDNA. Our method allows the percentage of doubly dsDNA in the total fluorescent dsDNA pool to be measured. In method, we excite the imperfectly labeled dsDNA sample in a focal of <1 fL with a laser beam and correlate the fluctuations of the signal to get the FCS autocorrelation curves; we express amplitudes of the autocorrelation function as a function of the DNA efficiency; we perform a comparative analysis of a dsDNA sample a reference dsDNA sample, which is prepared by increasing the total concentration c (c > 1) times by adding unlabeled ssDNA during the process. The method is flexible in that it allows for the of the reference sample and the c value can be adjusted as for a specific study. We express the precision of the method as a of the ssDNA labeling efficiency or the dsDNA double labeling The measurement precision can be controlled by changing the c value.
Journal: LASER PHYSICS LETTERS
Volume: 11
ID: ISI:000338982000021
Year: 2014
DOI: 10.1088/1612-2011/11/8/085602