Cells were washed with 5 ml ice-cold PBS prior to centrifugation for 5 min at 300 RCF, 4 °C. 800 μl of lysis buffer (50 mM Tris-HCl pH 7.4, 10 mM NaCl, 100 mM KCl 0.1 μM CaCl2, 0.5 mM MgCl2, 0.5 mM TCEP, EDTA-free complete protease tablet (Roche, 04639159001) was used to lyse the cell pellet. Lysates were sonicated for 3x 3 secs using a Microson ultrasonic cell disrupter XL (Misonix) on ice. Lysates were subjected to microcentrifugation at 4 °C for 12 min at 16000 RCF and the supernatants were transferred to fresh tubes.
Microson ultrasonic cell disrupter xl
Manufactured by Bioventus
Sourced in United States
The Microson ultrasonic cell disrupter XL is a laboratory equipment designed for the disruption of cells and tissue samples. It utilizes high-frequency sound waves to break down cellular structures, allowing for the extraction and isolation of cellular components, such as proteins, nucleic acids, and organelles.
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Lab products found in correlation
4 protocols using microson ultrasonic cell disrupter xl
1
Cell Lysis and Protein Extraction
2
Efficient dsRNA Extraction from Bacteria
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Sonication or heat (95°C) treatments were applied before isolating dsRNA from bacterial cells by conventional dsRNA purification process. One milliliter of the cell suspension (= 12.5 mL initial cell culture equivalent) was sonicated on ice using an ultrasonic cell disruptor (Microson Ultrasonic Cell Disrupter XL, Misonix, Wallkil, NY, USA) equipped with a microprobe. The sonication was performed at the strength of 5 to 6 W (outcome) on ice for 30 seconds 5 times with 30-second interval between sonications. The other 1 mL of cell suspension was heated at 95°C for 10 minutes with shaking at 350 r/min in ThermoMixer C (Eppendorf, Hauppauge, NY, USA). After sonication or heat treatment, dsRNA was extracted using the conventional method started with the 65°C incubation in the phenol/chloroform/isoamyl alcohol and quantified by NanoDrop 2000 spectrophotometer. The quality of the dsRNA was verified after separation by electrophoresis through a 1.2% agarose gel. To extract crude dsRNA, the cells lysed by sonication were centrifuged at 12,000 g for 30 minutes at 4°C. The supernatant was evaluated for dsRNA by gel electrophoresis and bacteria viability by spreading onto LB agar plates as described above.
3
Cell Lysis and Protein Extraction
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Cells were gently washed with 5 ml ice-cold PBS before collected and pelleted at 300 RCF for 5 min. The cell pellet was lysed in 800 µl of lysis buffer (50 mM Tris–HCl pH 7.4, 10 mM sodium chloride, 100 mM potassium chloride, 0.1 µM calcium chloride, 0.5 mM magnesium chloride, 0.5 mM TCEP, EDTA-free complete protease tablet (Roche, 04639159001)). The lysates were sonicated on ice for 3 × 3 s using a Microson ultrasonic cell disrupter XL (Misonix), with an output power of 0.06 Watts (RMS Watts). The lysates were clarified by microcentrifugation at 4°C for 12 min at 16 000 RCF. Supernatants were transferred to fresh tubes.
4
Cell Lysis and Protein Extraction
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Cells were gently washed with 5 ml ice-cold PBS before collected and pelleted at 300 RCF for 5 minutes. The cell pellet was lysed in 800 µl of lysis buffer (50 mM Tris-HCl pH 7.4, 10 mM sodium chloride, 100 mM potassium chloride, 0.1 µM calcium chloride, 0.5 mM magnesium chloride, 0.5 mM TCEP, EDTA-free complete protease tablet (Roche, 04639159001)). The lysates were sonicated on ice for 3x 3 seconds using a Microson ultrasonic cell disrupter XL (Misonix), with an output power of 0.06 Watts (RMS Watts). The lysates were clarified by microcentrifugation at 4 °C for 12 minutes at 16000 RCF. Supernatants were transferred to fresh tubes.
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