Reinhart Wesley F, Reifenberger Jeff G, Gupta Damini, Muralidhar Abhiram, Sheats Julian, Cao Han, Dorfman Kevin D
Department of Chemical Engineering and Materials Science, University of Minnesota-Twin Cities, 421 Washington Ave SE, Minneapolis, Minnesota 55455, USA.
BioNano Genomics, 9640 Towne Centre Dr., Ste. 100, San Diego, California 92121, USA.
J Chem Phys. 2015 Feb 14;142(6):064902. doi: 10.1063/1.4907552.
We obtained experimental extension data for barcoded E. coli genomic DNA molecules confined in nanochannels from 40 nm to 51 nm in width. The resulting data set consists of 1 627 779 measurements of the distance between fluorescent probes on 25 407 individual molecules. The probability density for the extension between labels is negatively skewed, and the magnitude of the skewness is relatively insensitive to the distance between labels. The two Odijk theories for DNA confinement bracket the mean extension and its variance, consistent with the scaling arguments underlying the theories. We also find that a harmonic approximation to the free energy, obtained directly from the probability density for the distance between barcode labels, leads to substantial quantitative error in the variance of the extension data. These results suggest that a theory for DNA confinement in such channels must account for the anharmonic nature of the free energy as a function of chain extension.
我们获得了宽度在40纳米至51纳米之间的纳米通道中受限的条形码大肠杆菌基因组DNA分子的实验延伸数据。所得数据集包含对25407个单个分子上荧光探针之间距离的1627779次测量。标签之间延伸的概率密度呈负偏态,且偏态的大小对标签之间的距离相对不敏感。两种用于DNA受限的奥戴克理论界定了平均延伸及其方差,这与理论背后的标度论证一致。我们还发现,直接从条形码标签之间距离的概率密度获得的自由能的谐波近似,会导致延伸数据方差出现显著的定量误差。这些结果表明,此类通道中DNA受限的理论必须考虑自由能作为链延伸函数的非谐波性质。