Microseal b

Manufactured by Bio-Rad

Sourced in United States

Microseal B is a sealing film product designed for use with microplates and other lab equipment. It provides a tight seal to prevent sample evaporation and contamination during various laboratory procedures.

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Lab products found in correlation

Infinite f50, by Tecan (3 mentions) 96 well flat bottom plate, by BD (2 mentions) Mx3000p qpcr machine, by Agilent Technologies (2 mentions) Klustercaller software, by LGC (2 mentions) Thermomixer comfort, by Eppendorf (1 mentions) 384 well plate, by Bio-Rad (1 mentions) Facsmelody, by BD (1 mentions) Dna away, by Thermo Fisher Scientific (1 mentions) Fluostar omega, by BMG Labtech (1 mentions) Clariostar, by BMG Labtech (1 mentions) Puregrade, by Brandel Inc (1 mentions) Cfx96 touch real time pcr detection system, by Bio-Rad (1 mentions) Pcr primers, by Bio-Rad (1 mentions) Lightcycler 480 2, by Roche (1 mentions) Primer mix, by Merck Group (1 mentions) Premade kasp master mix, by LGC (1 mentions) Primers mix, by Merck Group (1 mentions) Mxpro software, by Agilent Technologies (1 mentions) C1000 touch thermal cycler, by Bio-Rad (1 mentions) Hard shell pcr plate, by Bio-Rad (1 mentions)

Spelling variants of this product

Microseal ‘B’ seal (16 citations) Microseal B film (12 citations) Microseal B adhesive sealer (11 citations) Microseal® ‘B’ PCR Plate Sealing Film (9 citations) Microseal ‘B’ Adhesive Seals (9 citations) Microseal ‘B’ adhesive sealing film (2 citations) Microseal B clear (2 citations) PCR Sealers tape (Microseal ‘B’ Film) (2 citations) Microseal B adhesive (2 citations) Microseal ‘B’ Adhessive Sealer (2 citations)

11 protocols using microseal b

Single-cell sorting of E. coli

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Escherichia coli K12 MG1655 (DSMZ 18039) was cultured in 1 mL of Luria Bertani (LB) broth at 30 °C and 750 rpm with the Thermomixer Comfort (Eppendorf, Hamburg, Germany) to the exponential growth phase (~4 h; OD600 of ~2.2–2.6). From this point forward, cells were processed in a UV-decontaminated ISO 4 cleanroom. Equipment and gloves were decontaminated with DNA AWAY (Thermo Fisher Scientific, Waltham, MA, USA). Consumables were UV treated for 1 hr in a Crosslinker and 1 × PBS was UV treated for 6 h in a 254 nm shortwave ultraviolet crosslinker at 0.12 Joules/I² (Analytik Jena GmbH, Jena, Germany).
A BD FACSMelody (Becton-Dickson, Franklin Lakes, NJ, USA), fitted with a 100 µM nozzle and equipped with a 488 nm laser for excitation was used to sort single cells. Cells were first diluted to approximately 10⁶ cells mL⁻¹ with sterile 1X PBS to ensure an event rate of <1000 events/s. Gates were defined on side-scatter (cell complexity) and forward-scatter (cell-size). Cells were sorted in single-cell mode into 384-well plates (Bio-Rad, Hercules, CA, USA) containing no sorting buffer (i.e., dry sorting). Plates were sealed with Microseal B (Bio-Rad, Hercules, CA, USA) and stored at −80 °C.

Sobol M.S, & Kaster A.K. (2023). Back to Basics: A Simplified Improvement to Multiple Displacement Amplification for Microbial Single-Cell Genomics. International Journal of Molecular Sciences, 24(5), 4270.

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Fluorescence Measurements in Bulk

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Fluorescence measurements in bulk were conducted at 29°C in a plate reader (FLUOstar Omega or CLARIOstar, BMG Labtech, Germany). Fifteen microliters of samples were pipetted in a 384-well plate (pureGrade, BRAND, #781622), and the plate was sealed with an optically transparent film (Microseal “B”; Bio-Rad, Germany) and centrifuged at 700 relative centrifugal force (rcf) for 30 s.

Dupin A., Aufinger L., Styazhkin I., Rothfischer F., Kaufmann B.K., Schwarz S., Galensowske N., Clausen-Schaumann H, & Simmel F.C. (2022). Synthetic cell–based materials extract positional information from morphogen gradients. Science Advances, 8(14), eabl9228.

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Quantitative PCR Amplification and Detection

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qPCR reactions contained 1X Bio-Rad SsoFast Supermix (1725201, Bio-Rad), PCR primers (IDT) at 0.5 µM each, and were supplemented with nuclease-free water up to 10 µL. Each 96-well plate (thin-wall clear well, HSP9641, Bio-Rad) was sealed (Microseal B, MSB1001, Bio-Rad) and briefly spun in a Mini Plate Spinner Centrifuge (14-100-141, Fisher Scientific). Heating and real-time imaging were performed on the Bio-Rad CFX-96 Touch Real-Time PCR Detection System by heating to 95 °C for 5 min, cycling 40 times between 95 °C for 15 s, 60 °C for 15 s, and 72 °C for 20 s, and taking a melt-curve analysis. For the E. coli DNA dilution experiment, qPCR was run for 60 cycles. Fluorescence readings were taken at the end of each extension step. Quantification cycle (C_q) was determined when the software’s automated baseline corrected fluorescence reached 200 RFU.

Jue E., Witters D, & Ismagilov R.F. (2020). Two-phase wash to solve the ubiquitous contaminant-carryover problem in commercial nucleic-acid extraction kits. Scientific Reports, 10, 1940.

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Anaerobic Growth Assay of AIEC

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Starter cultures of AIEC in PYG were sub-cultured in reduced, sterile PYG containing 250mM or 1mM of 4-PBA or equivalent DMSO control. Growth assays were performed in sterile, clear, flat-bottom 96-well plates (Falcon), overlayed with sterile mineral oil, and sealed with an optically clear plate seal (Microseal ‘B’, BioRad). Growth was measured every 15 minutes by absorbance, OD₆₀₀, on a Tecan Infinite F50 microplate reader with shaking at 7.8 Hz between readings. Growth assays were conducted at 37°C in the anaerobic chamber.

Viladomiu M., Khounlotham M., Dogan B., Lima S.F., Elsaadi A., Cardakli E., Castellanos J.G., Ng C., Herzog J., Schoenborn A.A., Ellermann M., Liu B., Zhang S., Gulati A.S., Sartor R.B., Simpson K.W., Lipkin S.M, & Longman R.S. (2022). Agr2-associated ER stress promotes adherent-invasive E. coli dysbiosis and triggers CD103+ dendritic cell IL-23-dependent ileocolitis. Cell reports, 41(7), 111637.

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Differential Scanning Fluorimetry Assay for NEIL1

Cited 1 time

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The DSF assay was done according to the protocol previously described by Michel et al. [46 (link)]. In Brief, 0.2 μL triplicates in a 2:1 dilution of a 10 mM TH5487 solution and a DMSO control were spotted in White BioRad 384-well plates. Next, 9.8 μL of protein buffer, containing 25 mM Tris-acetate pH 7.5, 150 mM NaCl, 10% glycerol, 1 mM DTT, 5x SYPRO Orange, and 4 μM NEIL1 lacking 56 amino acids dispensable for activity at the disordered C-Terminus, were added to each well, yielding a final protein concentration of 3.92 μM and a final compound concentration ranging from 1.56 μM to 50 μM. The plates were sealed with BioRad MicroSeal ‘B’, centrifuged at 1000 rpm and subjected to a temperature gradient of 20 to 95 °C in a freshly started and cold Roche Light Cycler 480 II. The resulting fluorescence at 465–580 nm with an excitation wavelength of 465 nm was measured. Visual quality control of each graph was performed, and the resulting data were processed and imported to a GraphPad Prism template provided by Niesen et al. [47 (link)].

Hanna B.M., Michel M., Helleday T, & Mortusewicz O. (2021). NEIL1 and NEIL2 Are Recruited as Potential Backup for OGG1 upon OGG1 Depletion or Inhibition by TH5487. International Journal of Molecular Sciences, 22(9), 4542.

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Genotyping of F2 Populations using KASP Assay

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A subset of the positively identified SNP markers from the BSA were selected for genotyping the F₂ populations from the AC × PI 207262 and AC × G 2333 crosses. Genotyping was accomplished using kompetitive allele specific PCR (KASP) assays. The KASP primer sequences were designed using Primer3 [52 (link),53 (link)], and the KASP reactions were conducted following the manufacturer's instructions in 10 μL reactions with 5 μL of 2X premade KASP master mix (LGC, Middlesex, UK), 0.14 μL of primer mix (Sigma-Aldrich, St. Louis, USA), and 20–40 ng of genomic DNA. PCR parameters were performed as described by LGC on standard thermocycling machines using white semiskirted polypropylene 0.2 mL 96-well PCR plates (USA Scientific) and sealed with Microseal B (Bio-Rad, Hercules, CA, USA). After PCR amplification was completed, the PCR plates were scanned using the Mx3000P qPCR machine (Agilent, Santa Clara, CA), and allele calls for each genotype were obtained using KlusterCaller software (LGC, Middlesex, UK).

Gilio T.A., Hurtado-Gonzales O.P., Gonçalves-Vidigal M.C., Valentini G., Ferreira Elias J.C., Song Q, & Pastor-Corrales M.A. (2020). Fine mapping of an anthracnose-resistance locus in Andean common bean cultivar Amendoim Cavalo. PLoS ONE, 15(10), e0239763.

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KASP Genotyping of F2 Populations

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A subset of SNPs positively associated with Ur-3 found using BSA were selected for genotyping the F₂ population from Pinto 114 × Aurora using Kompetitive Allele Specific PCR (KASP) assays. Additional SNPs used for KASP genotyping were retrieved from SNP chip tables found in Song et al. (2015) (link). KASP primers were designed using the PrimerExpress software and KASP reactions were conducted following the manufacturer’s instructions. The 10 μl reaction contained 5 μl of 2× premade KASP master mix (LGC, Middlesex, UK), 0.14 μl of primers mix (Sigma-Aldrich, St. Louis, MO), and 20–40 ng of genomic DNA. PCR parameters were as described by LGC, on standard thermocycling machines, using white semiskirted polypropylene 0.2 ml 96-well PCR plates (USA Scientific), and sealed with MicrosealB (Bio-Rad, Hercules, CA). After PCR amplification was completed, PCR plates were scanned using the Mx3000P qPCR machine (Agilent, St. Clara, CA) and allele calls for each genotype were obtained using the MxPro software (Agilent) or the Klustercaller software (LGC).

Hurtado-Gonzales O.P., Valentini G., Gilio T.A., Martins A.M., Song Q, & Pastor-Corrales M.A. (2016). Fine Mapping of Ur-3, a Historically Important Rust Resistance Locus in Common Bean. G3: Genes|Genomes|Genetics, 7(2), 557-569.

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Anaerobic Growth Kinetics of AIEC

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Starter cultures of AIEC in PYG were subcultured in reduced media specified by experiment in a sterile, clear, flat-bottom 96 well plate (Falcon®), overlayed with sterile mineral oil, and sealed with an optically clear plate seal (Microseal® ‘B’, BioRad). Growth was measured every 15 minutes by absorbance, OD₆₀₀, on a Tecan Infinite F50 microplate reader with shaking at 7.8 Hz between readings. Growth assays were conducted at 37 °C in the anaerobic chamber.

Viladomiu M., Metz M.L., Lima S.F., Jin W.B., Chou L., Bank J.L., Guo C.J., Diehl G.E., Simpson K.W., Scherl E.J, & Longman R.S. (2021). Adherent-Invasive E. coli metabolism of propanediol in Crohn’s disease regulates phagocytes to drive intestinal inflammation. Cell host & microbe.

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Thermal Stability Screening of DNA Repair Enzymes

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White BioRad 384-well plates with
duplicates of each compound, DMSO, and positive control were prepared.
Next, 9.8 μL of protein buffer, containing 25 mM Tris-acetate
pH 7.5, 150 mM NaCl, 10% glycerol, 1 mM DTT, 5X SYPRO Orange, and
4 μM NEIL1, lacking 56 amino acids dispensable for activity
at the disordered C-Terminus, was added to each well, yielding a final
protein concentration of 3.92 μM and a final compound concentration
of 1 mM. Similarly, 4 μM OGG1 was screened in 25 mM Tris-acetate
pH 7.5, 50 mM CaCl₂, 10% glycerol, 1 mM DTT, 5X SYPRO Orange.
The plates were sealed with BioRad MicroSeal ‘B’ and
subjected to a temperature gradient from 20 to 95 °C in a Roche
Light Cycler 480 II. The resulting fluorescence at 465–580
nm with an excitation wavelength of 465 nm was measured. Visual quality
control of each graph was performed, and the resulting data were processed
and imported to a GraphPad Prism template provided by Niesen et al.^{92 (link)} For optimization and DNA experiments not concerning
the fragment screens, black BioRad Hard-Shell PCR 96-well thin-wall
plates were used in a BioRad C1000 Touch Thermal Cycler with 510 nm
excitation wavelength and measurement at 465 nm. NEIL2, APE1, and
TDG were prepared in a protein buffer containing 4 μm protein,
25 mM Tris-acetate pH 7.5, 150 mM NaCl, 10% glycerol, 1 mM DTT, and
5X SYPRO Orange.

Michel M., Visnes T., Homan E.J., Seashore-Ludlow B., Hedenström M., Wiita E., Vallin K., Paulin C.B., Zhang J., Wallner O., Scobie M., Schmidt A., Jenmalm-Jensen A., Warpman Berglund U, & Helleday T. (2019). Computational and Experimental Druggability Assessment of Human DNA Glycosylases. ACS Omega, 4(7), 11642-11656.

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Potato DNA Quantification via Asymmetric PCR

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PCR was performed using 60 ng of potato DNA in 10 µl reaction volume [1× PCR buffer, 2.5 mM MgCl₂, 0.2 mM each dNTP, 0.02 µM forward primer, 0.2 µM reverse primer, 0.2 µM probe (Integrated DNA Technologies)] and 0.5 unit of Taq polymerase (Invitrogen). PCR was performed on a PTC-100 thermocycler (MJ Research Inc.) in 96-well plates. The program used was the following: 2 min at 94 °C, followed by 55 cycles of 40 s at 94 °C, 40 s at annealing temperature (Ta), and 45 s at 72 °C, then 5 min at 72 °C as a final extension step. Finally, the reaction was cooled to 25 °C. PCR products were evaluated on a 1% agarose gel electrophoresis. The asymmetric PCR was performed in Hard-Shell PCR plates, 96 wells, thin walled (Bio-Rad, cat. HSP9665) with the addition of 1× LC Green (BioFire Defense). The reaction mixture was overlaid with 15 µl of mineral oil and the plates were covered with adhesive film Microseal B (Bio-Rad, cat. MSB1001). An extra step of 30 s at 94 °C was added to the PCR program to allow the formation of the hybrid between the probe and the strand in excess.

Herrera M.D., Vidalon L.J., Montenegro J.D., Riccio C., Guzman F., Bartolini I, & Ghislain M. (2018). Molecular and genetic characterization of the Ryadg locus on chromosome XI from Andigena potatoes conferring extreme resistance to potato virus Y. TAG. Theoretical and Applied Genetics. Theoretische Und Angewandte Genetik, 131(9), 1925-1938.

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