Human iPSC-OVs were manually selected and differentiated to d30, fixed in 1% formaldehyde for 10 min at room temperature, washed, and lysed in Pierce IP lysis buffer (Thermo Fisher) supplemented with 40 μl/ml protease cocktail inhibitor P8340 (Sigma Aldrich). Cleared lysate was sheared in a Q700 ultrasonic processor (Qsonica) equipped with a cup horn, and shearing was monitored using 1% agarose gel electrophoresis. 10% volume was reserved for input and the remainder was incubated with 2 μg sheep anti-VSX2 antibody (Exalpha) overnight at 4°C with rocking. Immunoprecipitates were collected on protein-G conjugated Dynabeads (Invitrogen), washed 5x with sterile PBS, and eluted in 10 mM Tris/1 mM EDTA pH 8 + 1% SDS. Three volumes of 1% SDS, 0.1M NaHCO3, and 200 mM NaCl were added to input and IP samples and crosslinks were reversed by incubation at 65°C for 4 hours. DNA was extracted with phenol:chloroform:isoamyl alcohol, ethanol precipitated, and quantified with the Qubit high sensitivity double stranded DNA kit (Thermo Fisher). 8–10 ng of DNA were prepared for deep sequencing with either the Illumina ChIP-Seq DNA or the TruSeq ChIP Sample Preparation Kit (Illumina) and quantified with a Qubit fluorometer. All samples were loaded at a final concentration of 8 pM and sequenced on the Illumina HiSeq 2500.
Q700 ultrasonic processor
Manufactured by Qsonica
Sourced in United States
The Q700 ultrasonic processor is a laboratory equipment designed to perform high-powered ultrasonic processing. It is capable of generating high-frequency sound waves to disrupt samples or facilitate various processes in a controlled laboratory environment.
Automatically generated - may contain errors
Lab products found in correlation
Zetasizer nano ultra, by Malvern Panalytical (2 mentions)
Sc250exp speedvac concentrator, by Thermo Fisher Scientific (2 mentions)
Rvt5105 refrigerated vapor trap, by Thermo Fisher Scientific (2 mentions)
Pierce ip lysis buffer, by Thermo Fisher Scientific (2 mentions)
Truseq chip sample preparation kit, by Illumina (1 mentions)
Dynabead, by Thermo Fisher Scientific (1 mentions)
Hiseq 2500, by Illumina (1 mentions)
208 s morgagni, by Thermo Fisher Scientific (1 mentions)
Magellan 400l, by Thermo Fisher Scientific (1 mentions)
Nanobrook 90plus, by Brookhaven Instruments (1 mentions)
7 protocols using q700 ultrasonic processor
1
Chromatin Immunoprecipitation of VSX2 in Human iPSC-Derived Organoids
2
Characterization of Electrochemical Inverse Emulsions
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Electrochemical experiments were performed using a CHI 750 potentiostat (CHI Instruments, Austin, TX, USA) with 3-electrode systems in a faradaic cage. Inverse emulsions were prepared using a Q700 ultrasonic processor (Qsonica, Newtown, CT, USA). Dynamic light scattering (DLS) was used to measure particle size using a NanoBrook 90Plus particle size analyzer (Brookhaven Instruments Corporation, Holtsville, NY, USA). Two types of reference electrodes were used in two different solvents. In aqueous solutions, a Ag/AgCl reference electrode (0.197 V vs. NHE) containing saturated KCl was used, whereas, in organic solvents, a Ag/Ag+ reference electrode (0.541 V vs. NHE) was prepared using a silver wire and a membrane containing 10 mM AgNO3 and 100 mM TBAPF6 in ACN.
3
Characterization of Graphene Particles
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Stock suspensions of GPs at a concentration of 250 μg/mL were prepared by dispersing powder in 0.02% sodium cholate and sonicated by a sonic probe (QSonica, Q700 ultrasonic processor, LLC, Newtown, CT, USA) for 30 min with a 65% amplitude. The hydrodynamic diameter and zeta potential of GPs in suspension further diluted in sterile water or culture medium (10% FBS) were determined using Zetasizer Nano-Ultra (Malvern Panalytical Ltd., Malvern, UK). The absence of biological contamination was confirmed using a cell-based assay as previously stated by Svadlakova et al. [25 (link)]. The stock solution was diluted in different concentrations and exposed to cell culture media with 10% FBS.
4
Characterization of Nanoparticle Suspensions
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Stock suspensions of GP at a concentration 250 µg/mL were prepared by dispersing powders in 0.02% sodium cholate, followed by sonication using a sonic probe (QSonica, Q700 ultrasonic processor with a 1/4" microtip probe) for 30 min with 65% of amplitude. The average shape and size were assessed by transmission electron microscopy (TEM, Philips 208 S Morgagni, FEI) at an accelerating voltage of 80 kV and by scanning electron microscopy (SEM, Magellan 400L, FEI). The hydrodynamic diameters (DH) of GP were determined using Zetasizer Nano-Ultra (Malvern Panalytical Ltd, Malvern, UK). Measurements were provided in several dilution of stock solutions in Milli-Q water and cell culture media containing of 10% FBS.
Stock suspension of MWCNT at a concentration 500 µg/mL was prepared by dispersing powder in 0.02% sodium cholate and sonicating using QSonica (Melville, NY, USA), Q700 5 min with 55% of amplitude. The average length and diameters of all used materials were assessed by TEM at an accelerating voltage of 80 kV and by SEM.
Stock suspension of MWCNT at a concentration 500 µg/mL was prepared by dispersing powder in 0.02% sodium cholate and sonicating using QSonica (Melville, NY, USA), Q700 5 min with 55% of amplitude. The average length and diameters of all used materials were assessed by TEM at an accelerating voltage of 80 kV and by SEM.
5
Electrochemical Characterization of Emulsions
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Electrochemical experiments were performed using a 760E potentiostat (CH Instruments, Austin, TX, USA) with a three-electrode cell placed in a Faraday cage. A 25 μm Pt-UME was used as a working electrode. Ag/AgCl (in saturated KCl) and Ag/Ag+, 10 mM AgNO3 (in acetonitrile) reference electrodes were chosen for the aqueous and organic solvent phases, respectively, whereas a Pt wire (1 mm diameter) was used as the counter electrode. To analyze the particle size, dynamic light scattering (DLS, NanoBrook 90Plus, Brookhaven Instruments Corporation, Holtsville, NY, USA) was used. A microtip-equipped horn sonicator (Q700 ultrasonic processor, Qsonica, Newtown, CT, USA) was used to create the water-in-oil emulsions. Before all the experiments, the working electrode was rinsed with acetone, ethanol, and water.
6
Metabolite Extraction from Nematode Samples
7
Metabolite Extraction from Nematode Samples
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Approximately 10,000 mixed-staged worms in 1.5mL microfuge tubes were lyophilized for 18-24 hrs using a VirTis BenchTop 4K Freeze Dryer. After the addition of two stainless steel grinding balls and 1mL of 80% methanol, samples were sonicated for 5 min (2 sec on/off pulse cycle at 90 A) using a Qsonica Q700 Ultrasonic Processor with a water bath cup horn adaptor (Model 431C2). Following sonication, microfuge tubes were centrifuged at 10,000 RCF for 5 min in an Eppendorf 5417R centrifuge. 800μL of the resulting supernatant was transferred to a clean 4mL glass vial, and 800μL of fresh methanol added to the sample. The sample was sonicated and centrifuged as described, and the resulting supernatant was transferred to the same receiver vial and concentrated to dryness in an SC250EXP Speedvac Concentrator coupled to an RVT5105 Refrigerated Vapor Trap (Thermo Scientific). The resulting powder was suspended in 120μL of 100% methanol, followed by vigorous vortex and brief sonication. This solution was transferred to a clean microfuge tube and subjected to centrifugation at 20,000 RCF for 10 min in an Eppendorf 5417R centrifuge to remove precipitate. The resulting supernatant was transferred to an HPLC vial and analyzed by HPLC-MS.
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