After 2 weeks of treatment, blood samples were collected for analysis on the 15th day. Fish blood was collected through a caudal vein or artery using a syringe. The whole blood was collected in a 1.5 mL microcentrifuge tube (Eppendorf) and centrifuged at 1006×g and 4°C for 10 min. The upper layer containing serum was collected. Testosterone and estradiol levels in the serum were measured using the sandwich - enzyme-linked immunosorbent assay method (Bioassay Technology Laboratory, Shanghai, China), according to the manufacturer’s protocols. The absorbance was measured using a microplate reader at 450 nm. Finally, the testosterone and estradiol levels were determined using a standard curve.
1.5 ml microcentrifuge tube
Manufactured by Eppendorf
Sourced in Germany
The 1.5 mL microcentrifuge tube is a laboratory equipment designed for sample preparation and storage. It has a capacity of 1.5 milliliters and is commonly used in various scientific applications.
Automatically generated - may contain errors
Lab products found in correlation
Dna enzymatic labelling kit, by Agilent Technologies (2 mentions)
C elegans 4x44k microarray, by Agilent Technologies (2 mentions)
G2539a scanner, by Agilent Technologies (2 mentions)
Ge wash buffer 1, by Agilent Technologies (2 mentions)
Ge wash buffer 2, by Agilent Technologies (2 mentions)
Nanodrop nd 1000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Feature extraction software 10, by Agilent Technologies (2 mentions)
Hybridization buffer and blocking agent, by Agilent Technologies (2 mentions)
Transplex complete whole transcriptome amplification kit, by Merck Group (2 mentions)
Miseq platform, by Illumina (1 mentions)
Hemocytometer, by Hausser Scientific (1 mentions)
Benchtop centrifuge, by Eppendorf (1 mentions)
0.5 ml microcentrifuge tube, by Eppendorf (1 mentions)
12 protocols using 1.5 ml microcentrifuge tube
1
Fish Blood Analysis of Hormones
2
Saliva DNA Extraction Protocol
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For the isolation of the total genomic DNA, standard chloroform-phenol protocol was applied with slight modification. 1 ml saliva sample was placed in a 1.5 ml microcentrifuge tube (Eppendorf AG, Hamburg, Germany) and centrifuged to collect cell pellet. In the cells, 0.5 ml of lysis buffer (containing NaCl, ethylenediaminetetraacetic acid (EDTA), sodium lauriate sulfate and NP40), and 0.01% proteinase-K were added, mixed gently and incubated at 55°C for 45–60 minutes for complete lysis of cells. After incubation, 250 µl of chloroform and phenol were added, mixed gently and centrifuged at 10,000 rpm for 5 min. The supernatant was then transferred to a new microcentrifuge tube and repeat chloroform-phenol step. The DNA was precipitated from the supernatant with two volumes of iso-propenol (ice-cold). The DNA pellet was washed with 70% ethanol, dried, dissolved in Tris-EDTA buffer (containing 10 mM Tris-HCl, 1 mM EDTA, pH 7.6) and stored at −20°C. The concentration of extracted DNA was adjusted to 1 μg/µl.
3
Poxvirus Genome Sequencing Protocol
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Tissue from individual pox lesions was aseptically dissected and mechanically homogenized in lysis buffer using disposable tissue grinder pestles and transferred into a 1.5 mL microcentrifuge tube (Eppendorf). Virion enrichment and DNA extraction from the sample was performed according to the protocol described by Sarker et al.25 (link),52 (link). DNA libraries were prepared according to published protocol53 (link) using the Illumina Nextera XT DNA Library Prep V3 Kit starting with one ng of total genomic DNA (gDNA) as measured by Qubit (Invitrogen) and sequenced on the Illumina MiSeq platform.
4
Modular Centrifugal-Force Driven Micronozzle
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The centrifugal-force driven modular micronozzle device was composed of a modular micronozzle, a micronozzle supporter, and a collection tube. These materials were obtained commercially. To fabricate the modular micronozzle, a syringe needle (KOVAX-NEEDLE®, Korea Vaccine Co., Ltd., Seoul, Korea) was inserted into the bottom of a 1.5-mL microcentrifuge tube (Eppendorf AG, Hamburg, Germany). A hole was bored by heating up the 23-gauge needle. Subsequently, connections were sealed with epoxy resin to prevent liquid leakage. Subsequently, the micronozzles were inserted into the square holes (i.e., 1 cm × 1 cm) on the tube cap as a micronozzle supporter that can bore a hole, such that the micronozzle would pass through. Finally, a 50-mL centrifuge tube (Becton Dickinson, Cowley, Oxford, UK), as a collection tube, was assembled with screws. Unlike other microfluidic methods, surface treatment was not required in this assembly.
5
Serum Protein Digestion and Labeling
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To 10 μL of sera, 990 μL of 1× PBS, pH 8.5 was added in a 1.5 mL microcentrifuge tube (Eppendorf, USA). When mentioned, samples were reduced with 5 mM dithiothreitol for 30 min at 60°C, then alkylated with 10 mM 2‐iodoacetamide for 60 min at 25°C in the dark. The digestion was performed in 8‐well 350 μL PCR sample strips (FisherScientific, USA). To each well containing 30 μL of diluted serum, 5 μL of labeled IS peptide mix, 5 μL 0.6% RSF, and 5 μL of 1 mg/mL trypsin was added followed with digestion at 37°C for 3 h. To quench the digestion and degrade the acid labile RSF detergent, 3 μL of 0.5N HCl was added, followed by a 30‐s mixing on a benchtop vortexer at 200 rpm, and incubation for 1 h at 37°C.
6
Propagating Nosema ceranae Spores in Honey Bees
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Nosema ceranae spores were extracted from the Nosema-infected colony of A. florea obtained from the local area of the Chon Buri Province, Thailand. We first propagated the spores by group-feeding A. mellifera workers (5 × 107 spores for 50 bees) collected from colonies located at the honey bee research unit of Burapha University, Chon Buri, Thailand. We kept these caged bees at 34 ± 2 °C (Memmert IPP 260, Schwabach, Germany) with 50–55% RH (Barigo-8861, Schwenningen, Germany) for 14 days. Midguts of propagation honey bees were removed and transferred to a 1.5 mL microcentrifuge tube (Eppendorf, Hamburg, Germany) containing 100 µL distilled water. Spores were extracted based on the standard method described by Fries et al. (2013) [33 (link)] and Naree et al. (2021) [26 (link)]. They were counted under a light microscope (Olympus CX50, Shinjuku, Tokyo, Japan) using a hemocytometer (Hausser Scientific, Horsham, PA, USA) following the procedure described by Cantwell (1970) [34 ]. Spores were further centrifuged and then resuspended in 50% (w/v) sucrose solution to standardize the concentration to 5 × 105 spores per µL. Spores were further identified as N. ceranae using Polymerase Chain Reaction (PCR) technique (Figure S1 ).
7
Transcriptome Analysis of C. elegans
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Worms were collected as young adults 46 h after a 1 h synchronization by collecting hatching L1 larvae as described above. Single worms were then washed, collected in a 1.5 ml microcentrifuge tube (Eppendorf) and snap-frozen in liquid nitrogen. Total RNA was extracted using an RNAClean XP kit (Agentcourt) following the manufacturer’s instructions and amplified using the TransPlex Complete Whole Transcriptome Amplification Kit (Sigma Aldrich) to generate cDNA. Cy3-labeled cDNA was prepared from 500 ng of double-stranded cDNA using the DNA Enzymatic Labelling Kit (Agilent) according to the manufacturer’s instructions, followed by purification with a 30 kDa column (Amicon). Dye incorporation and cDNA yield were checked with the NanoDrop ND-1000 Spectrophotometer (ThermoScientific). cDNA was mixed with hybridization buffer and blocking agent (Agilent) and incubated at 95 °C for 3 min before cooling on ice. cDNA was then hybridized to a custom C. elegans 4x44K microarray (Agilent) for 40 h at 65 °C. Microarrays were washed 1 min at room temperature with GE wash buffer 1 (Agilent) and 1 min at 37 °C with GE wash buffer 2 (Agilent), then dried immediately by brief centrifugation. Microarrays were scanned on a G2539A scanner (Agilent) at 5 µm resolution and 100 % PMT. Probe intensities were extracted and their quality was assessed with the Feature Extraction software 10.7 (Agilent).
8
Oral Microbiome Sampling Protocol
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All participants were asked to withhold their teeth cleaning and avoid eating and drinking at least 1 h before sample collection. Two oral samples were collected from each individual: saliva and occlusal plaque. Participants were instructed to rinse with water for the saliva collection to remove all saliva from the mouth. Then, 2–3 mL of unstimulated saliva was collected by drooling/spitting directly into a 50 mL Falcon conical sterile tube (Fisher Scientific, Pittsburg PA, USA) kept on ice during the collection. The posterior occlusal plaque was collected using one Pikster™ (Erskine oral care, Marian del Rey, CA, USA) per each quadrant, and all 4 quadrants were pooled (four-quadrant sample) into a sterile 1.5 mL micro-centrifuge tube (Eppendorf, AG, Hamburg, Germany) containing glycerol to a final concentration of 25%. All the collected samples, either saliva or plaque, were frozen immediately and stored at −80 °C.
9
Hemolymph Isolation from Challenged Larvae
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Twelve larvae were challenged with 10 μL of approximately 107 CFU/mL of C. perfringens JBCNJ055 washed culture and 12 larvae injected with 10 μL of 0.1% peptone water as controls. All larvae were incubated at 37°C simultaneously for 72 h. At 24 h intervals post challenge, three larvae from each group were placed into separate sterile 15 mL centrifuge tubes and submerged in ice for 15 min to immobilize. Haemolymph was extracted by removing the posterior two segments and bleeding into a sterile pre-chilled 1.5 mL micro centrifuge tube (Eppendorf, United Kingdom). 25 μL of extracted haemolymph was transferred into 75 μL of 50 mM PBS (pH 6.5) (Gibco, United Kingdom) then briefly vortexed and centrifuged to pellet the cells at 11,180 ×g for 10 min at 4°C. The supernate was transferred into sterile 1.5 mL tubes. Samples were processed within 15 min of extraction to avoid rapid melanisation.
10
Serum Sample Preparation Protocol
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Blood samples were centrifuged at 3000× g for 5 min, after which the harvested serum sample was transferred into a 1.5 mL microcentrifuge tube (Eppendorf). Then, the serum samples were stored at -20°C temperatures until when used.
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