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Sputum Liquification Buffer
Simple and efficient liquification
Sputum liquification to allow for the isolation of cells
Liquify sputum prior to nucleic acid isolation
Norgen’s Sputum Liquification Buffer is designed for the liquification of viscous sputum samples prior to isolation of cells for microscopy or nucleic acids for molecular testing. The homogenization of sputum samples is often desirable, particularly when there is a requirement for centrifugation to pellet cells or prior to the isolation of nucleic acids. The Sputum Liquification Buffer contains the mucolytic agent dithiothreitol which acts by reducing the disulfide bonds of the mucus proteins, thereby decreasing sample viscosity and allowing for easy handling and processing. Genomic DNA or RNA can then be isolated from the homogenized sample as Norgen’s Sputum Liquification Buffer is both DNAse and RNAse-free.
Figure 1. Isolation of Genomic DNA from Sputum Samples Liquefied using Norgen's Sputum Liquification Buffer. Expectorated sputum samples (0.2 mL) were collected and an equal volume of Norgen's Sputum Liquification Buffer (final concentration = 50 µg/mL dithiothreitol) was added to each sample. Samples were then incubated at 37°C with intermittent mixing to allow the samples to be completely homogenized. Genomic DNA was then isolated from the homogenized sputum samples using Norgen's Sputum DNA Isolation Kit (Cat# 46200) according to the kit protocol. For each 100 µL DNA elution, 10 µL was loaded into the wells of a 1X TAE, 1% agarose gel, run at 150 volts for 30 minutes. The gel was then visualized via ethidium bromide staining and photographed.
SHELF LIFE AND HANDLING
The Sputum Liquification Buffer should be stored at 15-30°C. When stored properly, the reagent should remain stable for at least two years. Diluted solution should be used as soon as possible, with unused portions stored at 4°C.