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Typhoon trio fluorescence scanner

Manufactured by GE Healthcare
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The Typhoon TRIO fluorescence scanner is a versatile imaging system designed for a wide range of life science applications. It utilizes fluorescence detection technology to capture high-quality images of gels, blots, and microplates. The Typhoon TRIO features multiple excitation lasers and emission filters, allowing for the detection of a variety of fluorescent dyes and labels.

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Lab products found in correlation

Phusion hot start 2 polymerase, by Thermo Fisher Scientific (1 mentions) Fluorchem gel documentation system, by Bio-Techne (1 mentions) Mini protean, by Bio-Rad (1 mentions) Powerpac hc, by Bio-Rad (1 mentions)

Close spelling variants of this product

Typhoon Trio (260 citations) Typhoon scanner (254 citations) Typhoon Trio scanner (84 citations) Typhoon fluorescence scanner (13 citations) Trio scanner (4 citations) Scanners (2 citations) Typhoon Trio high performance fluorescence scanner (2 citations)

5 protocols using typhoon trio fluorescence scanner

1

Quantifying dsDNA using Gel Electrophoresis

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Gel electrophoresis was used to quantify the dsDNA content on each paper layer after ITP of the dsDNA ladder. Gel electrophoresis was chosen for two reasons. First, most common dsDNA stains, such as SYBR gold or EtBr, exhibit a high background on cellulose paper, and this makes it difficult to visualize and quantify the amount of dsDNA. Second, the dsDNA ladder is comprised of twelve dsDNA components having lengths ranging from 100 to 1517 bp. Gel electrophoresis can separate them and provides quantitative information for each component of the ladder.
The gel electrophoresis analyses were conducted as follows. First, each fold of the paper was cut off, dried, and then inserted into a 1.3% agarose gel containing 10 µg/mL EtBr (Figure S2). Control samples were prepared by drying 1.0 µL of the 500 µg/mL dsDNA ladder stock solution in the paper zone. Gel electrophoresis was run using 1× TAE (containing 40.0 mM tris, 20.0 mM acetic acid, and 1.0 mM EDTA) buffer for 50 min at 100 V (Lambda LLS9120 DC Power Supply). A Typhoon Trio fluorescence scanner (GE Healthcare, Piscataway, NJ) was used to image the gel, followed by ImageJ software analysis.
Li X., Luo L, & Crooks R.M. (2015). Low-Voltage Paper Isotachophoresis Device for DNA Focusing. Lab on a chip, 15(20), 4090-4098.
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2

Intasome-Mediated Strand Transfer Assay

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Reactions were carried out using intasomes purified by size exclusion chromatography. A standard reaction contained 1.25 μg of intasome and 5 μg of nucleosome in 300 μl of 240 mM NaCl, 5 mM MgCl2, 1 mM DTT, 4 μM ZnCl2, 25 mM BisTris propane-HCl, pH 7.45. Strand transfer was allowed to proceed for 15 min at 37 °C, and the reaction was stopped by addition of 0.5% SDS and 25 mM EDTA. DNA products, deproteinized by digestion with 30 μg proteinase K at 37 °C for 1 h and ethanol precipitation, were separated in 4-12% TBE PAGE gels. Fluorescein-labeled DNA was detected using a Typhoon TRIO fluorescence scanner (GE Healthcare); non-labeled DNA was visualized by staining with GelRed. Strand transfer assays using naked supercoiled plasmid target DNA was done according to published procedures2 (link).
Maskell D.P., Renault L., Serrao E., Lesbats P., Matadeen R., Hare S., Lindemann D., Engelman A.N., Costa A, & Cherepanov P. (2015). Structural basis for retroviral integration into nucleosomes. Nature, 523(7560), 366-369.
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3

Chlamydomonas Gene Expression Using Venus

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The open reading frames (ORFs) of Chlamydomonas genes were expressed in frame with Venus from the PsaD promoter using the pLM005 vector. ORFs were amplified from genomic DNA using Phusion Hotstart II polymerase (Thermo Fisher Scientific, www.thermofisher.com) with the respective oligos in Table S1. HpaI‐cut pLM005 vector and PCR products were gel purified and assembled by Gibson assembly (Gibson et al., 2009). Due to the large gene length of HLA3, it was cloned in two fragments then assembled in the pLM005 vector by Gibson assembly. The pLM005 vector contains the AphVIII gene for paromomycin resistance in Chlamydomonas and ampicillin resistance for bacterial selection. All construct junctions were verified by Sanger sequencing. Constructs were transformed into Chlamydomonas by electroporation as in Zhang et al. (2014). Briefly, 250 μL of 2 × 108 cells/mL was transformed with 14.5 ng/kbp of EcoRV‐cut plasmid at 16 °C. Cells were spread on 86 mL TAP agar plates containing paromomycin (20 μg/mL) and kept in low light (~10 μmol photons/m2/s) until colonies were ~2–3 mm in diameter. Plates were screened for fluorescent colonies using a Typhoon TRIO fluorescence scanner (GE Healthcare, www.gelifesciences.com) with excitation/emission wavelengths 532 nm/520–555 nm for Venus and 633 nm/630–670 nm for chlorophyll autofluorescence.
Atkinson N., Feike D., Mackinder L.C., Meyer M.T., Griffiths H., Jonikas M.C., Smith A.M, & McCormick A.J. (2015). Introducing an algal carbon‐concentrating mechanism into higher plants: location and incorporation of key components. Plant Biotechnology Journal, 14(5), 1302-1315.
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4

Intasome-Mediated Strand Transfer Assay

Cited 1 time
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Reactions were carried out using intasomes purified by size exclusion chromatography. A standard reaction contained 1.25 μg of intasome and 5 μg of nucleosome in 300 μl of 240 mM NaCl, 5 mM MgCl2, 1 mM DTT, 4 μM ZnCl2, 25 mM BisTris propane-HCl, pH 7.45. Strand transfer was allowed to proceed for 15 min at 37 °C, and the reaction was stopped by addition of 0.5% SDS and 25 mM EDTA. DNA products, deproteinized by digestion with 30 μg proteinase K at 37 °C for 1 h and ethanol precipitation, were separated in 4-12% TBE PAGE gels. Fluorescein-labeled DNA was detected using a Typhoon TRIO fluorescence scanner (GE Healthcare); non-labeled DNA was visualized by staining with GelRed. Strand transfer assays using naked supercoiled plasmid target DNA was done according to published procedures2 (link).
Maskell D.P., Renault L., Serrao E., Lesbats P., Matadeen R., Hare S., Lindemann D., Engelman A.N., Costa A, & Cherepanov P. (2015). Structural basis for retroviral integration into nucleosomes. Nature, 523(7560), 366-369.
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5

Quantifying RNA-Peptide Interactions via EMSA

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Electrophoretic mobility shift assays (EMSAs) were performed using a Bio-Rad Mini-Protean gel system paired with a direct current (DC) power source (PowerPac™ HC, Bio-Rad). Typically, 6 μL solutions containing ds RNA (c = 3 µM) and N-terminally fluorescein-isothiocyanate-labeled peptide (c = 6 µM) were incubated for 1 hour at 4 °C in a binding buffer of 10 mM Tris (pH = 8.0), 0.5 mM EDTA, 12.5 % glycerol, and 150 mM KCl. After incubation, bound RNA was resolved from peptide-free RNA using 10 % non-denaturing polyacrylamide gels at 120 V in running buffer (25 mM Tris (pH = 8.0), 200 mM glycine, 20 mM NaCl) at 4 °C for 1 hour. Visualization of fluorescent gel-bound species was conducted using a Typhoon Trio + fluorescence scanner (GE Healthcare Life Sciences). For nucleic acid visualization gels were stained using 2 µL of SYBR™ gold nucleic acid gel dye (Molecular Probes) in 20 mL of 1×TAE buffer (40 mM Tris, 1 mM EDTA, 20 mM acetic acid) for 45 minutes at RT before being visualized using a FluorChem™ gel documentation system (Alpha Innotech).
Kuepper A., McLoughlin N.M., Neubacher S., Camps E.C., Nithin C., Mukherjee S., Bethge L., Bujnicki J.M., Brock R., Heinrichs S., & Grossmann T.N. (2020). Constrained Peptides Mimic a Viral Suppressor of RNA Silencing.
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