Cells were mated on ME agar by crossing h+ and h− strains and adding sterile water. The plates were incubated at 26 °C for 3–5 days for growth and sporulation to form zygotic asci, which were picked for tetrad dissection. After sporulation, a loop of crossed cells was streak on fresh YES or EMM plates to pick individual asci and place each spore in a grid matrix using the microdissection microscope Singer MSM System Series 400 (Singer Instruments, Somerset, UK). The four spores of the zygotic ascus were placed each at a unique position in a straight line using the micromanipulator. The plates were then incubated at 26 °C. After colony formation, the plates were replicated onto YES and more importantly, selective plates of YES with an antibiotic or EMM without specific supplements using filter paper (GE Healthcare Life Sciences, Little Chalfont, UK). When the spores grow after incubation at 26 °C, the plates are photographed using a ChemiDoc (Bio-Rad Laboratories Ltd., Watford, UK) for scoring and following genetic patterns. The desired genetic backgrounds for each colony was confirmed using PCR analysis. The viability of colonies on each plate was noted and total viable colonies were counted to calculate percentage viability and to exam the possibility of synthetic lethality by using the chi-square test.
Filter paper
Manufactured by GE Healthcare
Sourced in United States
Filter paper is a specialized absorbent material used in laboratory settings. It is designed to separate solid particles from liquids or gases by filtering them out. Filter paper is made from cellulose fibers and is available in various grades, pore sizes, and shapes to accommodate different filtration requirements. The primary function of filter paper is to facilitate the filtration process, enabling the efficient separation of mixtures and the purification of solutions.
Automatically generated - may contain errors
Lab products found in correlation
Veco grids, by Nisshin EM (3 mentions)
Ht7700 transmission electron microscope, by Hitachi (2 mentions)
Chemidoc, by Bio-Rad (1 mentions)
H 7700 transmission electron microscope, by Hitachi (1 mentions)
Mbt biotarget 96, by Bruker (1 mentions)
Microflex smart instrument, by Bruker (1 mentions)
Filter paper, by Thermo Fisher Scientific (1 mentions)
Tecnai spirit, by Thermo Fisher Scientific (1 mentions)
Carbon coated copper grid, by Electron Microscopy Sciences (1 mentions)
Rotary evaporator, by Büchi (1 mentions)
9 protocols using filter paper
1
Tetrad Analysis of Fission Yeast
2
Exosome Visualization via Negative Staining
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A 6-μL aliquot of exosomes was absorbed onto glow-discharged 300-mesh heavy-duty carbon-coated Cu grids (Veco grids; Nisshin EM, Tokyo, Japan) for 2 min and the excess was blotted on filter paper (Whatman; GE Healthcare, Piscataway, NJ, USA). The grids were then washed twice with MilliQ water and negatively stained with 2% uranyl acetate. Data were collected using an H7700 transmission electron microscope (Hitachi, Tokyo, Japan) operating at 80 kV and 10,000× magnification.
3
Visualizing Exosome Nanoparticles via TEM
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A 6-μL aliquot of the fractions containing exosomes was absorbed onto glow-discharged 300-mesh heavy-duty carboncoated Cu grids (Veco grids, Nisshin EM, Tokyo, Japan) for 2 min and the excess was blotted on filter paper (Whatman, GE Healthcare, Piscataway, NJ, USA). The grids were then washed twice with MilliQ water and negatively stained with 2% uranyl acetate. Data were collected using an HT7700 transmission electron microscope (Hitachi, Tokyo, Japan) operating at 80 kV and 10,000× magnification.
4
Virus Imaging by Negative Staining TEM
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A 6-μl aliquot of UV-irradiated virus was absorbed onto glow-discharged 300-mesh heavy-duty carbon-coated Cu grids (Veco grids; Nisshin EM, Tokyo, Japan) for 2 min, and the excess was blotted onto filter paper (Whatman; GE Healthcare, Piscataway, NJ). Grids were then washed twice with Milli-Q water and negatively stained with 2% phosphotungstic acid. Data were collected using an HT7700 transmission electron microscope (Hitachi, Tokyo, Japan) operating at 100 kV electrons and magnification of ×30,000.
5
Rapid Antimicrobial Susceptibility Testing
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DOT-MGA was performed as previously described (Idelevich et al., 2018a (link)). Briefly, microdroplets containing 3 μl of antibiotic solution and 3 μl of bacterial suspension (final inoculum approximately 5 × 105 CFU/ml) were pipetted onto an MBT Biotarget 96 (Bruker Daltonik, Bremen, Germany). Sterility and growth controls were spotted on a second target. Targets were incubated for 3 or 4 h at 35 ± 1°C using a plastic transport box (Bruker Daltonik) serving as a humidity chamber in order to prevent the microdroplets from evaporating. After incubation, the remaining liquid was removed from the spots using filter paper, (size 37 × 100 mm, GE Healthcare GmbH, Freiburg im Breisgau, Germany). After overlaying the spots with 1 μl of α-cyano-4-hydroxycinnamic acid matrix including internal standard (MBT MASTeR prototype kit, Bruker Daltonik), MALDI-TOF MS spectra were acquired on a microflex smart instrument (Bruker Daltonik). The method was performed in triplicate.
6
Virus Sample Integrity Verification
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The integrity of all purified virus samples was confirmed using transmission electron microscopy. For each sample, 2 µL of purified virus was applied to carbon-coated copper grids (Electron Microscopy Sciences, Hatfield, PA, USA) for 2 min followed by wicking of excess buffer with filter paper (GE Healthcare Life Sciences). Grids were washed with 10 µL of filtered water, excess liquid wicked off and then stained with 10 µL 1% uranyl acetate for 10 s. Stain was removed with filter paper and grids were imaged on a 120 keV Tecnai Spirit (Thermo Fisher Scientific).
7
Dried Blood Spot Sampling for gDNA Extraction
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Finger prick blood DBS Preparation. Finger prick blood was collected using a lancet to puncture the fingertip. Once a full small drop of blood is formed it was lightly touched to the center of the circle on the filter paper (GE Health Care Life Science, Catalog no. 10534612) to form valid DBS. These were collected from study participants during health checkup organized at government schools, and at the center of villages. Cards were dried for 2 hours in velcro rack and packed in sealed ziplock bag with 1–2gm desiccant sachet (
Figure 1 ). Only valid DBS samples were used for gDNA extraction determined by the blood sample completely saturating each circle on the card, and not overlapping or merging with other blood circles. Prepared DBS samples were transported to the laboratory at the Centre for Cancer Epidemiology, Tata Memorial Centre, where they were stored in a -80°C refrigerator.
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8
Immunocapture-based Bacterial DNA Extraction
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For detection, the samples were treated according to our previously developed immunocapture procedure [3 (link), 31 ]. Five milligrams of IMPs and 2 mL of each sample were added into different tubes followed by cultivation at 37°C for 30 min under stirring (150 rpm). Bacteria-bead complexes were separated by magnetic decantation, washed three times with 0.01 mol/L sodium phosphate buffer (PBS, pH 7.4) and resuspended in 200 μL TE buffer (10mM Tris-HCl, 1mM EDTA, pH = 8.0). Genomic DNA was released by boiling the solution for 10 min. After centrifugation at 6,000 ×g for 5 min (4°C), 100 μL of the supernatant was collected as template. Aliquots (50 μL) of the supernatant was purified by the filter paper (Whatman, General Electric Co., Fairfield,Conn., U.S. state) to remove the PCR inhibitor. The total volume of purified DNA was stored at -20°C and subjected to real-time PCR for detection. As a control, the other part of template was directly used for the real-time PCR amplification.
9
Extraction and Analysis of Siberian Ginseng Root Compounds
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Dried roots of SO were collected from China in November 2020 and purchased from a commercial market (Samhong, Gyunggi-do, Republic of Korea) in South Korea. Plant identification was confirmed by Professor Dae Sik Jang of Kyung Hee University (Seoul, Republic of Korea). The SO roots were dried and crushed into a powder. Powdered SO roots (50 g) were extracted with 500 mL of 95% ethanol for 72 h at room temperature on an orbital shaker and filtered through filter paper (GE Healthcare, Chicago, IL, USA). A rotary evaporator was used to evaporate the solvent in the SOEE under reduced pressure (Buchi, Flawil, Switzerland) to eliminate the solvent. The chromatogram of SOEE and standard compounds (ZG1, ZG2, and gallic acid) (Figure S1 ), chemical analysis information (Tables S1–S3 ), and quantification of each compound (ZG1, ZG2, and gallic acid) (Table S4 ) are shown in the Supplementary Materials .
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