Using a Bullet Blender Gold homogenizer (Next Advance, Averill Park, NY, USA), we homogenized whole ectoparasites in Navy Eppendorf bead tubes (Next Advance) containing 400 μL brain–heart infusion broth (CDC, Atlanta, GA, USA). We extracted DNA from the homogenates by using the KingFisher Flex Purification System and the associated MagMAX Pathogen RNA/DNA Kit (both ThermoFisher, Waltham, MA, USA) according to the manufacturer’s protocols. Detection of Bartonella DNA in ectoparasite samples was performed by nested PCR for gltA (20 (link)) because of low concentrations of DNA and by conventional PCR for ITS (21 (link)), followed by sequencing and sequence analysis of amplicons.
Bullet blender gold homogenizer
The Bullet Blender Gold homogenizer is a laboratory equipment designed for the rapid and efficient homogenization of samples. It is capable of grinding, mixing, and pulverizing a variety of materials with its high-speed blades. The device is engineered to provide consistent and reproducible results for various applications that require efficient sample preparation.
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7 protocols using bullet blender gold homogenizer
Ectoparasites Identification and Bartonella Detection
Using a Bullet Blender Gold homogenizer (Next Advance, Averill Park, NY, USA), we homogenized whole ectoparasites in Navy Eppendorf bead tubes (Next Advance) containing 400 μL brain–heart infusion broth (CDC, Atlanta, GA, USA). We extracted DNA from the homogenates by using the KingFisher Flex Purification System and the associated MagMAX Pathogen RNA/DNA Kit (both ThermoFisher, Waltham, MA, USA) according to the manufacturer’s protocols. Detection of Bartonella DNA in ectoparasite samples was performed by nested PCR for gltA (20 (link)) because of low concentrations of DNA and by conventional PCR for ITS (21 (link)), followed by sequencing and sequence analysis of amplicons.
Tissue Analysis of Murine Placenta
Bartonella Isolation from Rodent Hearts
Measuring Heat Stress Protein Synthesis
De novo protein synthesis was measured by adding 20 μL of 14C protein hydrolysate (Amersham CFB25, 50 μCi/mL) to 2 mL cell suspension for 1 h before lysis. After brief centrifugation, cells were resuspended in 250 mM NaOH and disrupted using a Bullet Blender Gold homogenizer (Next Advance, Inc., Averill Park, NY, USA) in the presence of zirconium oxide beads (0.5 mm) at speed 8 for 3 min at 4°C. The homogenate was centrifuged (1000 xg, 4°C, 3 min) and proteins were precipitated on ice by mixing samples with an additional 1/4 sample volume of 50% (v/v) aqueous TCA for 20 min. After centrifugation (10,000 xg, 4°C, 10 min), pellets were washed twice with 1 mL −20°C acetone and solubilized in 1x protein loading buffer for 3 h. Proteins were separated on 10% SDS-PAGE and prepared for fluorography as in [44 (link)].
Muscle RNA Extraction and RT-qPCR Analysis
Liver Biopsy Sampling for Bariatric Research
Quantitative Trehalose Analysis in Cell Lysates
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