For rapid mechanical lysis of microbial cells from low biomass and low inhibitor containing swabs, paper or filter paper for direct application to PCR for metagenomic analysis. This is not a DNA isolation kit.
Figure 1. PCoA plot displaying the distribution of samples color coded by cleaning regimen. DNA was released from swabs collected from gym equipment using the UltraClean®-htp 96 Well Swab DNA Kit. Each dot represents a different sample, and the three different colors represent three different types of cleaning regimens: spray cleaner (blue dots); cleaning wipes (red dots); and no regular cleaning (green dots). The grouping of the dots show qualitatively, that how the equipment is cleaned may influence the composition of the samples. Further investigation of the clustering trends shown in plots like these can determine if factors like cleaning play a significant role in microbial community structure.*
Figure 2. DNA purified with the UltraClean®-htp 96 Well Swab DNA Kit is ready to use in qPCR. Pellets of overnight E. coli culture were collected on sterile polyester swabs which were then processed in the UltraClean®-htp 96 Well Swab DNA Kit to release DNA in a final volume of 500 µl. 1 µl and a 1:10 dilution of solution containing released DNA were used directly for 16s qPCR with Bact/Pro primers, while the remaining solution was purified on a Thermo Scientific KingFisher® Duo using MO BIO's SwiftMag® Magnetic Bead Technology. Following purification, DNA was eluted into a final volume of 50 µl, a 10 fold concentration over the initial released DNA. 1 µl and a 1:10 dilution of this solution was used for 16s qPCR as described above. Cq values of the initial 1 µl of released DNA correlated with the 1:10 dilution of the purified DNA, indicating that no DNA was lost between the release and purification. Additionally, the results show that, for samples without inhibitors, DNA released using the UltraClean®-htp 96 Well Swab DNA Kit is pure enough for qPCR.
Figure 3. Taxa summary demonstrating which microbes are present at a given time. DNA was released from swab samples of hand weights in the gym using the UltraClean®-htp 96 Well Swab DNA Kit. A specific sample was tracked over the course of three days at the phylum level. Phylums examanined include Actinobacteria (purple), Bacteroidetes (blue), Cyanobacteria (brown), Firmicutes (green), Proteobacteria (pink) and “Other” bacteria (yellow).*
Figure 4. Plot of beta diversity within gym benches over a period of two days. DNA was released from swabs collected from gym benches using the UltraClean®-htp 96 Well Swab DNA Kit. Unifrac distances were used to determine, at a certain time, how different that time point is from a base time point. Data is plotted as collection time (minutes) away from time zero to determine the change in diversity over time.*
*Data courtesy of Mariah Wood1, 2, Jarrad Hampton-Marcell1, and Jack Gilbert1, Argonne National Laboratory1, Argonne, IL and Weinberg College of Arts and Sciences, Northwestern University, Evanston, IL2.
|Starting Amount||Single swab, piece of paper or filter paper small enough to submerge in a 500 µl volume|
|Equipment Required||96 well plate shaker and Plate adapter set|
|Solution SW1||211 ml|