Norgen Biotek offers kits in different standardized formats. You can view and compare kit components in the Components Table in the Product Overview section. For more information on format types, please visit our Format Information page.
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Sputum DNA Isolation Kit
Fast and easy processing using a spin column format
DNA can be isolated and detected from as little as 100 µL of sputum
Effective removal of PCR inhibitors
This kit constitutes an all-in-one system for the rapid isolation of DNA from sputum samples. It allows for the isolation of bacterial or eukaryotic DNA from the sputum samples using a convenient spin column format. The kit includes all the reagents needed to process sputum DNA including a sputum liquifier to reduce viscosity and facilitate DNA isolation. The protocol can be completed in 30 minutes. Purified DNA is of the highest quality and free from inhibitors, and can be used in sensitive downstream applications including PCR and qPCR.
A sputum specimen is the name given to the mucus which is expectorated from the lower airways. High quality sputum samples should contain very little saliva, as this may contaminate the sputum sample with oral bacteria. Sputum samples are typically evaluated to look for infections such as Moraxella catarrhalis, Mycobacterium tuberculosis, Streptococcus pneumoniae and Haemophilus influenza. Other pathogens can be detected in sputum as well, including HIV. In addition, sputum samples are useful in the detection of lung cancer or to evaluate chronic inflammation.
Figure 1. Consistent Isolation of Sputum DNA. Sputum samples were collected from 16 donors using the collection devices from Norgen's Sputum DNA Collection, Preservation and Isolation Kit (Cat. RU46100), and the DNA was subsequently isolated using Norgen's Sputum DNA Isolation Kit. Ten µL of the purified DNA from each sample was then run on a 1X TAE, 1% agarose gel for visual analyses. DNA was successfully isolated from all 16 of the samples and was of a high integrity. It is important to note that the yield of DNA in a typical sputum sample will vary greatly and is highly dependent on the health status of the donor. The DNA yields obtained using Norgen's kit were equal to or exceeding those of what is stated in the literature.
Figure 2. High Yield of Sputum DNA. Sixteen sputum samples were collected using Norgen's Sputum DNA Collection, Preservation and Isolation Kit (Cat. RU46100) and stored at room temperature for 1 day. DNA was subsequently purified using Norgen's Sputum DNA Isolation Kit, and the DNA yield was determined using the NanoVue Plus (GE Healthcare). The total average sputum DNA yield from 1.0 mL of preserved sputum of the 16 samples processed using Norgen's kit was 75.25 µg (indicated by the red line in the graph). Again it is important to stress that the yield of DNA obtained from each sample will vary from donor to donor as it relies heavily on the health status of the donor at the time the sample was taken.
Figure 3. High Quality of Sputum DNA. Sixteen different sputum samples were collected using Norgen's Sputum DNA Collection, Preservation and Isolation Kit (Cat. RU46100) and stored at room temperature for 1 day. DNA was subsequently purified using Norgen's Sputum DNA Isolation Kit, and the DNA yield was determined using a NanoVue Plus (GE Healthcare). The average A260nm:A280nm ratio (indicated by red line in graph) obtained for each of the samples using Norgen's kit was 2.05 indicating the DNA is of vary high quality.
* Average DNA yield will vary depending on the health status of the donor
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.
Norgen’s Sputum DNA Isolation Kit contains ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.