Multitron standard

Manufactured by Infors

Sourced in Switzerland

The Multitron Standard is a high-performance incubator shaker designed for a wide range of cell culture and microbiology applications. It features precise temperature control, orbital shaking, and a compact design to accommodate multiple vessels simultaneously.

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Lab products found in correlation

Sllgx13nk, by Merck Group (1 mentions) Zeocin, by InvivoGen (1 mentions) Nocodazole, by Merck Group (1 mentions) Aquatron, by Infors (1 mentions) Genejet rna purification kit, by Thermo Fisher Scientific (1 mentions) Tryptic soy broth (tsb), by Merck Group (1 mentions) Soluble starch, by Merck Group (1 mentions) Ecotron, by Infors (1 mentions)

Spelling variants of this product

Multitron (21 citations) HT Multitron Standard (9 citations) Multitron Standard shaker-incubator (3 citations) HT Multitron Standard incubator (3 citations) Multitron Standard orbital shaker (2 citations) Multitron Standard shaker (2 citations) Multitron Standard shaking incubator (2 citations) Multitron standard incubator (2 citations)

7 protocols using multitron standard

Screening and Quantification of 3-HP in P. pastoris

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P. pastoris strains were inoculated into 50 mL falcon tubes containing 5 mL of YPG (1% yeast extract, 2% peptone and 1% v/v glycerol) supplemented with 100 μg mL⁻¹ zeocin (InvivoGen, CA, United States). The cells were grown overnight at 30°C and 200 rpm in an incubator shaker Multitron Standard (Infors HT, Bottmingen, Switzerland) with a 2.5 cm orbit. 50 μL of overnight-grown cultures were used to inoculate each well of a 24 deep-well plate containing 2 mL of Buffered Minimal Glycerol (BMG) medium, containing 100 mM potassium phosphate buffer pH 6, 1.34% yeast nitrogen base (YNB), 1% v/v glycerol, and 0.4 mg L⁻¹ biotin. The cultures were incubated at 28°C and 220 rpm in the same incubator shaker using a platform with a slope of 20° to improve the aeration. The cultures were grown for 48 h to ensure the full consumption of the substrate. Thereafter, culture samples were centrifuged at 12,000 g for 5 min and filtered through a 0.20 µm pore size syringe filter (SLLGX13NK, Merck Millipore, CA, United States). The 3-HP was quantified using HPLC.
The parental clones PpHP2 and PpHP6 were screened in triplicates. Six to eight clones were screened in triplicates for the strains generated by single-homology integration (PpHP7, PpHP8, and PpHP9). One single clone was screened in triplicates for the strains generated using CRISPR-Cas9.

Fina A., Heux S., Albiol J, & Ferrer P. (2022). Combining Metabolic Engineering and Multiplexed Screening Methods for 3-Hydroxypropionic Acid Production in Pichia pastoris. Frontiers in Bioengineering and Biotechnology, 10, 942304.

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Yeast Cell Synchronization Techniques

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All yeast strains were derived from W303 and grown in rich medium (YEP) supplemented with 2% glucose (YPD) at 25°C unless otherwise stated. Cultures were agitated at 200 rpm (Multitron Standard, Infors HT). Strain numbers and relevant genotypes of the strains used are listed in the Key resources table, Supplementary file 4. To arrest the cells in G1, α-factor was added to a final concentration of 2 mg/L/h, every 30 min for 2.5 hr. Release was achieved by filtration wherein cells were captured on 1.2 μm filtration paper (Whatman GE Healthcare), washed with 1 L YPD, and resuspended in the appropriate fresh media. To arrest the cells in G2, nocodazole (Sigma) was added to the fresh media to a final concentration of 10 μg/mL and cells were incubated until the synchronisation was achieved (>95% large-budded cells). To inactivate temperature-sensitive alleles, fresh media were pre-warmed prior to filtration (Aquatron, Infors HT). To produce cells deficient in Pds5 using the AID system, cells were arrested with α-factor as described above. 30 min prior to release, auxin was added to 5 mM final concentration. Cells were then filtered as described above and released into YPD medium containing 5 mM auxin.

Petela N.J., Gonzalez Llamazares A., Dixon S., Hu B., Lee B.G., Metson J., Seo H., Ferrer-Harding A., Voulgaris M., Gligoris T., Collier J., Oh B.H., Löwe J, & Nasmyth K.A. (2021). Folding of cohesin’s coiled coil is important for Scc2/4-induced association with chromosomes. eLife, 10, e67268.

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Bacterial Growth Monitoring and RNA Isolation

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Bacterial cultures were grown in the 1.0x and 0.5x Tryptic Soy Broth (Merck Millipore, Darmstadt, Germany). For Arthrobacter sp., Bacillus sp., Pseudomonas sp., and Rhodococcus sp., inoculation (1% vol/vol) was performed using fresh bacterial cultures grown on Tryptic Soy Agar plates at 30°C. Inocula (OD₆₀₀ = 2) were prepared in the respective sterile growth medium. Bacterial cultures were grown in an orbital shaker Multitron Standard (Infors AG, Bottmingen-Basel, Switzerland) at 30°C, 180 rpm. Growth of the culture was monitored by measuring optical density at 600 nm. For Streptomyces sp., the growth curve was constructed according to Manteca et al. (2005) (link) using a diphenylamine colorimetric method (Zhao et al., 2013 (link)). Sampling of all the strains was performed in the middle of the exponential growth phase, at the transition phase, and at the beginning of the stationary growth phase. RNA isolation was done using the GeneJET RNA Purification Kit (Thermo Fisher Scientific) according to the manufacturer’s instructions. The integrity of RNA samples was analyzed using 1% agarose gel. RNA samples were stored at –20°C until use.

Bukelskis D., Dabkeviciene D., Lukoseviciute L., Bucelis A., Kriaučiūnas I., Lebedeva J, & Kuisiene N. (2019). Screening and Transcriptional Analysis of Polyketide Synthases and Non-ribosomal Peptide Synthetases in Bacterial Strains From Krubera–Voronja Cave. Frontiers in Microbiology, 10, 2149.

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Cultivation of Hfx. mediterranei for Bioactive Compound Production

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Hfx. mediterranei strain R-4 (ATCC33500) was used for all experiments. Cells were grown in a complex medium containing 12.5% (w/v) of inorganic salts [29 (link),88 (link)], 0.5% (v/w) yeast extract, and 0.5–2.5% (w/v) of a carbon source (glucose (BioChemica, CAS: 50-99-7), soluble starch (Merck, CAS:9005-84-9) or oxalacetic acid (Alfa Aesar CAS: 328-42-7)). pH was buffered using 30 mM Tris-HCl (pH 7.3). Growth conditions included 36.5 °C and shaking at 170 rpm (Infors HT Multitron Standard) based on the data reported by [24 (link)]. Cultures were incubated until three days had passed after the maximum of absorbance was reached (stationary phase of growth). Since BR is a secondary metabolite, the stationary phase is where the majority of BR is synthesized. Then, the cells were centrifuged at 7800 rpm for 30 min to remove the supernatant. The cells were washed twice with a 10% (w/v) inorganic salts solution. Growth-specific velocity (µ) Equation (1) and duplication time (Dt) Equation (2) were calculated for each growth condition as follows:

Giani M., Gervasi L., Loizzo M.R, & Martínez-Espinosa R.M. (2022). Carbon Source Influences Antioxidant, Antiglycemic, and Antilipidemic Activities of Haloferax mediterranei Carotenoid Extracts. Marine Drugs, 20(11), 659.

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Growth and Genetic Manipulation of Caulobacter crescentus

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C. crescentus strains were routinely grown at 30°C in liquid PYE medium in either an Infors HT Multitron Standard or Infors HT Ecotron rotating shaker set to 200 rpm, or on solid PYE agar. Prior to analysis, all liquid cultures were grown undisturbed in exponential phase at 30°C for 3 h to allow any aggregation from other sources to be dissolved or diluted. Heat shock experiments were performed by moving 100 ml flasks of 10 ml cultures growing at 30°C to a shaking incubator pre‐heated to the desired temperature. Media were supplemented with the following additives when indicated; xylose 0.3%, vanillate 500 M. Antibiotics were used at following concentration (concentration in liquid/solid media as µg/ml): kanamycin 5 (2.5 in the case of KJ956 and KJ957)/25, spectinomycin 25/400, gentamycin 0.625/5, tetracycline 1/2 and rifampicin 2.5/5. Experiments were generally performed in the absence of antibiotic when using strains in which the resistance cassette was integrated into the chromosome. E. coli strains for cloning purposes were grown in LB medium at 37°C, supplemented with antibiotics at following concentrations: kanamycin 30/50, spectinomycin 50/50, gentamycin 15/20, tetracycline 12/12 and rifampicin 25/50. Details on strain and plasmid construction are reported in Supporting Information and Supporting Information Table S1.

Schramm F.D., Schroeder K., Alvelid J., Testa I, & Jonas K. (2019). Growth‐driven displacement of protein aggregates along the cell length ensures partitioning to both daughter cells in Caulobacter crescentus. Molecular Microbiology, 111(6), 1430-1448.

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Yeast Growth Optimization in Buffered Minimal Medium

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Cultures were first grown overnight on 5 ml of YPG medium (1% yeast extract, 2% peptone and 1% v/v glycerol) supplemented with the appropriate antibiotic in 50‐ml Falcon tubes, at 30°C, and 180 rpm. The overnight cultures were then diluted to an optical density at 600 nm (OD₆₀₀) of 0.5 on 5 ml of YPG and grown for 8 h at the same cultivation conditions. Afterwards, these cultures were used to inoculate a 500‐ml baffled shake flask containing 50 ml of buffered minimal glycerol medium (BMG; 100 mM potassium phosphate buffer pH 6, 1.34% yeast nitrogen base (YNB), 0.4 mg l⁻¹ biotin and 1% v/v glycerol) at a starting OD₆₀₀ of 0.05. The shake flask cultures were grown at 30°C and 180 rpm in an incubator shaker Multitron Standard (Infors HT, Bottmingen, Switzerland) with a 2.5 cm orbit.

Fina A., Brêda G.C., Pérez‐Trujillo M., Freire D.M., Almeida R.V., Albiol J, & Ferrer P. (2021). Benchmarking recombinant Pichia pastoris for 3‐hydroxypropionic acid production from glycerol. Microbial Biotechnology, 14(4), 1671-1682.

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Biogas Production from Birch Biomass

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The biogas production of untreated, steam-exploded, and HL birch was studied in 500 ml batch bottles. In addition, control batch bottles were prepared using inoculum alone to correct for the endogenous biogas production. All the bottles received equal amount of inoculum and anaerobic medium. The inoculum-to-substrate ratio was 2:1 (based on VS basis) as suggested by Holliger et al. [24 (link)]. The substrates were added individually into the batch bottles, while the control bottles received water instead of substrate. The bottles were flushed with nitrogen gas for 5 min and sealed with septum and aluminum caps to maintain anaerobic conditions. All the experiments were conducted in triplicate inside a shaker (Multitron Standard, Infors HT, Switzerland) at mesophilic condition (37 °C, 90 rpm). The biogas experiment run for 39 days and terminated when the daily biogas rate on 3 consecutive days was below 1% [24 (link)].

Mulat D.G., Dibdiakova J, & Horn S.J. (2018). Microbial biogas production from hydrolysis lignin: insight into lignin structural changes. Biotechnology for Biofuels, 11, 61.

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