Phosphorimager screen

Manufactured by Kodak

Sourced in United States

The Phosphorimager screen is a device used for detecting and quantifying radioactive signals in biological samples. It functions by capturing the light emitted from phosphor-coated screens that have been exposed to radioactive materials, allowing for high-resolution imaging and analysis of these samples.

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Lab products found in correlation

Hybond, by Cytiva (2 mentions) Biomax mr film, by Kodak (2 mentions) Gradient sds page gel, by Bio-Rad (2 mentions) Dntps, by Thermo Fisher Scientific (1 mentions) Hybond xl nylon membrane, by GE Healthcare (1 mentions) Dynazyme ext dna polymerase, by Thermo Fisher Scientific (1 mentions) Dna engine peltier thermal cycler, by Bio-Rad (1 mentions) Personal fx phosphorimager, by Bio-Rad (1 mentions) Ultrahyb hybridization buffer, by Thermo Fisher Scientific (1 mentions) Fla 9000 phosphorimager, by Fujifilm (1 mentions) Hybond n membrane, by Cytiva (1 mentions) Nucleospin mirna kit, by Macherey-Nagel (1 mentions) Molecular imager fx, by Bio-Rad (1 mentions)

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Phosphorimager (4 citations)

6 protocols using phosphorimager screen

Genotyping DMPK (CTG)n Repeats

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Genomic DNA was isolated from muscle sections following standard procedures. To amplify expanded (CTG)n repeats a heat pulse extension PCR protocol was used [38 (link)], that allows the generation of DMPK (CTG)n amplicons of up to 1750 CTG repeats. 40 ng DNA was used in a PCR mixture containing 2.25 M betaine (Fluka, Sigma-Aldrich, Germany), 0.2 mM dNTPs (Invitrogen, Carlsbad, CA, USA), 1.33 units DyNAzyme EXT DNA Polymerase (Thermo Scientific, Waltham, MA, USA) and 250 nM of each forward (5´-GCCAGTTCACAACCGCTCCGAGCGTGGGTC-3´) and reverse (5´-ACGCTCCCCAGAGCAGGGCGTCATGC-3´) primers (Biolegio BV, Nijmegen, the Netherlands). Cycling conditions were kept as described [38 (link)], except that an annealing temperature of 66°C followed by ramping to 83°C (0.9°C/sec) was used. PCR was performed in a DNA engine Peltier Thermal Cycler (Bio-Rad, Hercules, CA, USA). The size of the expected DMPK amplicon was 324 bp long excluding the (CTG)n repeat.
PCR products were run on a 1% agarose gel and transferred to a Hybond-XL nylon membrane (Amersham, GE Healthcare). The blot was incubated with a ³²P-end-labeled (CAG)9 probe and after washing exposed to a phosphor imager screen (Kodak, Rochester, NY, USA) for 3 hours. Signal was developed using a Personal FX Phosphor Imager (Bio-Rad).

González-Barriga A., Kranzen J., Croes H.J., Bijl S., van den Broek W.J., van Kessel I.D., van Engelen B.G., van Deutekom J.C., Wieringa B., Mulders S.A, & Wansink D.G. (2015). Cell Membrane Integrity in Myotonic Dystrophy Type 1: Implications for Therapy. PLoS ONE, 10(3), e0121556.

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Northern Blot Analysis of H. pylori RNA

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Total RNA was extracted at different time points after rifampin addition (80 µg/ml) from 5-ml samples of H. pylori cultures at an OD₆₀₀ of 0.5 to 0.9 by using a NucleoSpin miRNA kit from Macherey-Nagel (4 (link)). For Northern blotting, 5 µg of RNA was separated on an agarose-formaldehyde gel and transferred to a Hybond N+ membrane (Amersham Biosciences, Inc.) overnight by passive transfer in 10× SSC (pH 7) buffer (1× SSC is 0.15 M NaCl plus 0.015 M sodium citrate). Transferred RNA was fixed to the membrane via UV irradiation for 2 min. The membrane was blocked for 45 min at 65°C with ULTRAhyb hybridization buffer (Ambion), then 5 µl of ³²P-labeled riboprobe was added and the membrane was further incubated overnight at the same temperature. After three washes for 10 min at 65°C with 2× SSC, 0.2% SDS, the membrane was exposed to a phosphorimager screen (Kodak) and scanned with an FLA-9000 phosphorimager (Fujifilm).

El Mortaji L., Aubert S., Galtier E., Schmitt C., Anger K., Redko Y., Quentin Y, & De Reuse H. (2018). The Sole DEAD-Box RNA Helicase of the Gastric Pathogen Helicobacter pylori Is Essential for Colonization. mBio, 9(2), e02071-17.

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Polyacrylamide Gel Electrophoresis Analysis

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The products of the cleavage reactions were separated on 8% denaturing polyacrylamide gels containing 8M urea. After electrophoresis, the gels were fixed in 10% (v/v) acetic acid, dried and subjected to autoradiography using a phosphorimager screen (Kodak), which was scanned with a Typhoon phosphorimager.

Sekibo D.A, & Fox K.R. (2017). The effects of DNA supercoiling on G-quadruplex formation. Nucleic Acids Research, 45(21), 12069-12079.

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35S-Methionine and 3H-Proline Tracing

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Male Wistar rats (30–45 day old) were used. Groups of ten animals were injected in both eyes using the same procedures described above with 1 daily injection over 3 days with 5 µl solution containing a mix of 3.3 µCi/µl 35S-methionine and 1µCi/µl 3H-proline. Animals were sacrificed after 20 days. Brains and eyes were harvested and flash frozen in cold isopentane. For some experiments, 20 µm frozen sagittal sections were collected on Superfrost® plus microscope slides using a cryostat. For other experiments, dissected fresh tissue from retina, LGN, frontal and visual cortices were collected and homogenized in RIPA buffer. Radiolabeled proteins in the lysates were quantified using a liquid scintillation counter from PerkinElmer™ or loaded onto 4–20% SDS/PAGE gradient gel (Bio-Rad). Proteins were separated by electrophoresis and transferred to nitrocellulose membranes. Radiolabeled tissue sections and blotted nitrocellulose membranes were exposed to PhosphorImager screens or Kodak® BioMax® MR films inside autoradiography cassettes for 1 week and imaged by PhosphorImager or developed in an X ray film developer.

Schiapparelli L.M., Sharma P., He H.Y., Li J., Shah S.H., McClatchy D.B., Ma Y., Liu H.H., Goldberg J.L., Yates JR I.I.I, & Cline H.T. (2022). Proteomic screen reveals diverse protein transport between connected neurons in the visual system. Cell reports, 38(4), 110287.

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Southern Blot DNA Detection Analysis

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After visualization, agarose gels were soaked in 1 M HCl for 30 min on a shaker at room temperature, rinsed with double distilled water, and soaked in alkaline transfer buffer (0.4 N NaOH, 1 M NaCl) for 15 min at room temperature on a shaker. The alkaline transfer buffer was replaced for another 15 min at room temperature. Gels were rinsed with double distilled water, and soaked in neutralizing buffer (1 M Tris (pH7.4), 1.5 M NaCl) for 15 min at room temperature on a shaker. BioDyne B0.45 nylon membranes were cut to size and soaked in 10x SSC buffer (1.5 M NaCl, 150 mM trisodium citrate, pH 7.0). DNA in the agarose gels was capillary transferred according to standard protocols. Membranes were prehybridized with 10 ml rapid hybrid buffer (Amersham Biosciences, Piscataway, NJ) for one hour at 75°C in a rotating oven. The probe (HSV-1 EcoRI restriction fragment JK, [5 (link), 6 (link)]) was labeled by random priming according to the manufacturer's instructions (Amersham Biosciences). The denatured probe was added to 5 ml of rapid hybrid buffer pre-warmed at 75°C, and hybridized in the rotating oven for 4 h at 75°C. Membranes were washed twice for 15 min with hybridization wash buffer 1 (75 mM NaCl, 7.5 mM sodium citrate, 0.5% SDS) at room temperature, rinsed with distilled water, and exposed to Kodak PhosphorImager screens. Signal was quantitated in a Bio-Rad FX molecular imager.

Hu M., Depledge D.P., Flores Cortes E., Breuer J, & Schang L.M. (2019). Chromatin dynamics and the transcriptional competence of HSV-1 genomes during lytic infections. PLoS Pathogens, 15(11), e1008076.

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35S-Methionine and 3H-Proline Tracing

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