Sizing single DNA molecules

Guo, Xuan-Hui; Huff, Edward J.; Schwartz, David C.

doi:10.1038/359783a0

Cited by 24 publications

(14 citation statements)

References 9 publications

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“…The length can quickly be measured simply by stretching and immobilizing the DNA on a surface [121]. This is useful for restriction mapping [122].…”

Section: Fractionation Of Dnamentioning

confidence: 99%

Micro- and nanofluidics for DNA analysis

Tegenfeldt

Prinz

Cao

et al. 2004

Analytical and Bioanalytical Chemistry

295

226

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Miniaturization to the micrometer and nanometer scale opens up the possibility to probe biology on a length scale where fundamental biological processes take place, such as the epigenetic and genetic control of single cells. To study single cells the necessary devices need to be integrated on a single chip; and, to access the relevant length scales, the devices need to be designed with feature sizes of a few nanometers up to several micrometers. We will give a few examples from the literature and from our own research in the field of miniaturized chip-based devices for DNA analysis, including dielectrophoresis for purification of DNA, artificial gel structures for rapid DNA separation, and nanofluidic channels for direct visualization of single DNA molecules.

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“…The length can quickly be measured simply by stretching and immobilizing the DNA on a surface [121]. This is useful for restriction mapping [122].…”

Section: Fractionation Of Dnamentioning

confidence: 99%

Micro- and nanofluidics for DNA analysis

Tegenfeldt

Prinz

Cao

et al. 2004

Analytical and Bioanalytical Chemistry

295

226

View full text Add to dashboard Cite

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“…One-half of a plug containing DNA was loaded directly into a gel well. The estimated sizing uncertainty by PFGE is ϳ10% (14). Lanes 1 and 5 are ladder PFG markers; lane 3 is Low Range PFG markers; lanes 2 and 4 are SmaI digestion of S. aureus DNA.…”

Section: Sizing Of Smai Digestion Of S Aureus Dna By Pfgementioning

confidence: 99%

Bacteria genome fingerprinting by flow cytometry

Huang¹,

Jett²,

Keller³

1999

Cytometry

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Background: A flow cytometry–based, ultrasensitive fluorescence detection technique has been developed that demonstrates unique advantages in the analysis of large DNA fragments over the currently most widely used technology, pulsed‐field gel electrophoresis (PFGE). The technique described herein is used to characterize the restriction fingerprints of the bacteria genome Staphylococcus aureus in this study. Methods: The isolation of the bacterial genomic DNA and the subsequent complete digestion by a restriction endonuclease were performed inside an agarose plug. Electroelution was used to move the DNA fragments out of the agarose plug into a solution containing low concentrations of spermine and spermidine, added to stabilize the large DNA fragments. DNA was stained with the bisintercalating dye thiazole orange homodimer (TOTO‐1) and subsequently introduced into our ultrasensitive flow cytometer from a capillary. Results: Individual DNA fragments up to 351 kbp were successfully handled and sized. The histograms of the burst sizes were generated from signals associated with individual fragments in <7 min with <2 pg of DNA. The sizing accuracy was better than 98%. In contrast, standard PFGE takes ≈20 h and requires ≈1 μg of DNA with a sizing accuracy of ≈90%. Conclusions: With the demonstrated success and advantages, our approach has the potential of being applied to fast, accurate bacteria species and strain identification. Cytometry 35:169–175, 1999. © 1999 Wiley‐Liss, Inc.

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“…The 9.0-kbp fragment was faintly visible on the gel but did not reproduce well in the photograph. The estimated sizing uncertainty by PFGE is ϳ10% (14).…”

Section: Sizing Of Smai Digestion Of S Aureus Dna By Pfgementioning

confidence: 99%