High-sensitivity microvolume UV absorption spectrometry for routine analysis of small-volume biological samples.
Biotechniques
; 70(5): 251-262, 2021 05.
Article
em En
| MEDLINE
| ID: mdl-33956504
ABSTRACT
Substantial improvement of microvolume UV absorption spectrometry in sensitivity, robustness and ease of operation was achieved for routine biological applications. A unique microtubing-based absorption cell (208 µm internal diameter) featuring enhanced light transmission with a liquid core waveguide technique provided dramatically enhanced absorption sensitivity, proportional to the extended path length (50 mm, from the typical 1 mm), while robust measurement performance was attained by implementation of preventive measures against bubble trapping along the light path. For pBR322 plasmid DNA, absorbance at 260 nm was reliably measurable down to 0.1 ng/µl with repeatability typically 2-3% RSD. The detection limit was 0.03 ng/µl dsDNA, far lower than the current state-of-the-art â¼1 ng/µl. Sample consumption for each measurement was 2.4 µl. Automated operation implemented for the first time in microvolume spectrophotometry facilitated the ease in handling with small-volume biological samples.
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Base de dados:
MEDLINE
Assunto principal:
Análise Espectral
/
DNA
Tipo de estudo:
Diagnostic_studies
Idioma:
En
Ano de publicação:
2021
Tipo de documento:
Article