DNA and protein were extracted from rat brain, heart, and small intestines. Proteomic coverage was compared for all tissue types.
There have been many studies of correlating gene and protein expression from single tissue samples1,2,3,4. Clinical samples
can be especially challenging as sample sourcing and quantity are often limited. This is certainly the case for tissue samples of
human origin that represent rare disorders. To make the most of rare tissues an ideal extraction method would be suitable for
extracting a multitude of analytes, including nucleic acids, proteins and small molecules. Furthermore, the method would be
capable of processing multiple samples simultaneously with high reproducibility.
The Bead Ruptor 96, Bead Mill Homogenizer is capable of rapid tissue dissruption via bead milling in 96 well plates, tubes or
cryomilling in stainless steel chambers. Herein, we evaluate the Bead Ruptor 96 for high-throughput tissue dissruption for
the purification of genomic DNA and proteins. Genomic DNA integrity is evaluated, and the protein repertoire is analyzed via
electrophoresis and bottom up semi-quantitative proteomics.
The main objective of this study was to compare the performance of Omni International’s Soil DNA Kit (26-013G/B) to that of Company M’s Soil DNA Isolation Kit in terms of DNA yield and quality.
Molecular analysis of soil DNA offers a direct solution for detecting microorganisms residing in soil and for studying microbial diversity. Isolation of DNA from soils is often challenging because of the presence of many contaminants, like humic acid, that can interfere with the extraction process and are inhibitory to several downstream applications. An ideal DNA extraction method should effectively eliminate inhibitory substances and maximize DNA yields. The main objective of this study was to compare the performance of Omni International’s Soil DNA Kit (26-013G/B) to that of Company M’s Soil DNA Isolation Kit in terms of DNA yield and quality, as well as amplification potential and sensitivity of detection using real-time PCR.
Demonstrate extraction of bacterial genomic DNA from soil and amplification of the 16S rDNA gene through PCR.
Soil microbiome research seeks to understand the diversity and abundance of microorganisms in various soil types as a function of environmental conditions. As a first step toward this goal, microbe nucleic acids must be extracted from the soil substrate. A major obstacle toward defining the soil microbiome is the ability to first culture these microorganisms to gain a better understanding of their ecology, diversity and species richness. Currently microbial cell culture media is selective and only certain isolates can be determined by this approach. As an alternative to the cell culture approach, a popular determination method is to directly extract and amplify microbial DNA from soil samples. Although this alternative method has given promising results, there remains hurdles that must be overcome. Most notably, soil is natively rich in substances such as humic acids, that inhibit polymerases and restriction enzymes making the amplification of DNA difficult. Fortunately, methods have been established to separate microbial DNA from inhibitors prior to DNA purification and amplification. One such approach is available in the Omni Soil DNA Purification kit and is demonstrated in this application note. Herein, we evaluate the Omni Soil DNA Purification Mini kit’s ability to extract DNA from Georgia red clay using the Bead Ruptor 24 Elite for mechanical dissociation of the soil/microbe samples prior to PCR inhibitor removal and DNA purification.