A simple approach for effective shearing and reliable concentration measurement of ultra-high-molecular-weight DNA

Pipetting and concentration measurement of viscous ultra-high-molecular-weight (UHMW) DNA samples is challenging and often highly imprecise. Effective guidelines for handling UHMW samples are missing in the field. Herein, a simple and low-cost workflow is presented that enables accurate pipetting an...

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Détails bibliographiques
Publié dans:BioTechniques. - 1991. - 71(2021), 2 vom: 03. Aug., Seite 439-444
Auteur principal: 'Giron' Koetsier, Paul A (Auteur)
Autres auteurs: Cantor, Eric J
Format: Article en ligne
Langue:English
Publié: 2021
Accès à la collection:BioTechniques
Sujets:Journal Article Research Support, Non-U.S. Gov't DNA extraction DNA shearing genomics glass beads nanodrop needle shearing quantification qubit plus... ultra-high-molecular-weight DNA DNA 9007-49-2
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520 |a Pipetting and concentration measurement of viscous ultra-high-molecular-weight (UHMW) DNA samples is challenging and often highly imprecise. Effective guidelines for handling UHMW samples are missing in the field. Herein, a simple and low-cost workflow is presented that enables accurate pipetting and reliable concentration measurement. Central to the workflow is the shearing of representative small aliquots of UHMW DNA samples to a fragment size <150 kb by vortexing them for 1 min with a glass bead in a round-bottomed 2-ml tube. Additionally, a solution is provided for accurate quantitation of high-molecular-weight DNA with fluorometric (Qubit [Thermo Fisher Scientific, MA, USA]) methods by using an appropriate genomic DNA standard, resulting in values that match spectrophotometric (Nanodrop [Thermo Fisher Scientific]) optical density readings 
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