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DNA was extracted from mosquitos to be used for PCR using the Bead Ruptor 96.
Mosquitos, specifically Aedes aegypti, are known to act as a host for vector borne diseases such as Yellow fever, Dengue
fever, and Zika (1). Most of these diseases are associated with tropical climates, but Aedes aegypti are found throughout
the Southeast United States, with a likely range from Virginia to Texas (2). To monitor the spread of vector borne diseases,
researchers typically sample mosquitos from the field and dissociate the organism to release the internal pathogens to
quantify pathogen load. Researchers require a fast and repeatable sample preparation technique to rapidly dissociate a large
number of mosquitos while maintaining a high degree of vector and pathogen lysis. Herein, we demonstrate dissociation
of Aedes aegypti using the Bead Ruptor 96 bead mill homogenizer for the purification of DNA and analysis by endpoint PCR.
Hops (as an analog to cannabis) were spiked with different cannabinoids, homogenized on the Prep 96, and analyzed using HPLC to determine recovery efficiency.
Cannabinoid quantification (“Potency Testing”) is the most common analytical method performed by cannabis producers and testing facilities. Producers are required to define the quantity of cannabinoids in cannabis-based products before release to the market. While there are regional variations in potency testing requirements and more than 60 cannabinoids present in cannabis, producers are typically required to define the quantity of the abundant and psychoactive cannabinoids, Tetrahydrocannabinol (THC), Tetrahydrocannabinolic Acid (THCA), Cannabidiol (CBD), Cannabidiolic Acid (CBDA), Cannabigerol (CBG), and Cannabinol (CBN) (Figure 1). 1-2
The most common method for cannabis potency testing is to mill the flower or edibles to create a homogenous mixture in the presence of an organic solvent such as methanol. Following centrifugation to pellet debris, the supernatant is further diluted prior to analysis by reverse phase HPLC or mass spectrometry. While the analytical methods are well defined and easily automated, the sample disaggregation process is low throughput and often tedious. Herein, we evaluate the utility of the Prep 96 automated homogenizer for the extraction of cannabinoids from a spiked cannabis analog.
RNA was extracted from multiple mouse tissues using mechanical and bead milling homogenization along with the Omni Tissue RNA kit. RNA yields and integrity were determined
The extraction and isolation of RNA is an integral part of downstream analyses such as RT-PCR, RT-qPCR, Northern blotting, and
cDNA library construction. The importance of using pure, intact RNA for these processes is well documented and a critical part
of downstream analysis success. It is well known that RNA is sensitive to degradation due mechanical shear, temperature, storage
conditions and freeze-thawing. Furthermore, RNA is highly susceptible to RNAse degradation following release of nucleases
during the tissue disaggregation process. Thus, proper sample handling during the homogenization process is crucial when
performing an RNA based assay.
The Omni Tissue RNA Purification Kit supports rapid RNA purification with reproducible RNA yields and integrity. Extraction
is based on bead based tissue disruption followed by RNA purification on silica spin columns. Herein, we demonstrate the
performance of the Omni Tissue RNA Purification Kit for RNA purification from multiple murine tissues following disaggregation
on the Bead Ruptor 12 Bead Mill Homogenizer. RNA integrity and yield was compared to tissues dissociated through cryomilling
in a mortar & pestle under liquid nitrogen.