Pseudo-stems, the basal section of the tiller, were harvested from each endophyte-infected plant. Samples were harvested into liquid nitrogen and then transferred to a freeze drier (Freezone Plus12, Labconco Corporation, Kansas City, MI, USA). Once lyophilized the samples were ground and homogenised with a bead mill (Omni Bead Ruptor 24, Omni International Inc., Kennesaw, GA, USA) in a 7 mL vial using a ¼ inch zirconium bead (30 seconds at 4.5 m/s). Sub-samples (50 mg) were extracted with 1 mL of the prepared extraction solvent (80% v/v methanol with 0.54 ng/mL ergotamine, 0.202 ng/ml festuclavine, and 1.7 ng/mL homoperamine as internal standards) in 2 mL plastic vials for 1 hour by end-over-end rotation (30 Hz) in the dark. After centrifuging (5000 g, 5 min), the supernatant was transferred to 2 mL amber HPLC vials for analysis. Along with the samples, duplicate reference samples of AR37 (for quantifying the epoxyjanthitrems) were similarly extracted and analysed.
Freezone plus 12
Manufactured by Labconco
Sourced in United States
The FreeZone Plus 12 is a freeze dryer designed for sample preparation. It features a 12-liter chamber and accommodates up to 6 sample shelves. The system is capable of reaching temperatures as low as -105°C and a vacuum level of 0.012 mbar. The FreeZone Plus 12 is intended for use in a variety of laboratory applications.
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Lab products found in correlation
5 protocols using freezone plus 12
1
Endophyte-infected Tiller Metabolite Extraction
2
Synthesis and Characterization of Carbon Nanodots
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Carbon nanodots were synthesized based on our published microwave-assisted method [9 (link),11 (link),12 (link)]. The samples were diluted with 10.0 mL of DDI water and then dialyzed against 5.0 L of DDI water using a 500–1000 Da MWCO (Spectra Por Float-A-Lyzer G2, 10 mL, G235063; MilliporeSigma; Darmstadt, Germany). The sample was dialyzed over three days under a gentle stir with a new DDI water replacement every 24 h. The resultant clear, brown-orange aqueous solution was lyophilized (Labconco FreeZone Plus 12; Kansas City, MO, USA) to produce the dried carbon nanodots. The Cary® Eclipse™ Fluorescence Spectrophotometer was used for characterizing the photoluminescence of carbon nanodots.
3
Polyelectrolyte Complex Formation and Properties
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To study the influence of the PEI:SPA ratio and acid treatment on the PECs’ formation and properties, different solutions were prepared with acidification (noted as “AC”) and without acidification. The details are summarized in Table 1 . All solutions were immediately frozen for 24 h before being freeze-dried at 0.01 mbar with the collector temperature set at −80 °C using a Labconco FreeZone Plus 12 (Labconco, Kansas City, MO, USA) to analyze the PEC’s properties.
4
Chromatographic Isolation of NA-enriched SPC
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A chromatography column was used with 250 mL reservoir with an internal diameter of 20.0 mm and a length of 305 mm. The isolation of the NA-enriched SPC was conducted using column chromatography according to the method by Vikbjerg, et al. [38 (link)] with slight modification. The fractions of SPC+PC were collected, and the solvent was removed by rotary evaporation. The purified SPC+PC was pre-frozen at −80 °C for 12 h and subsequently lyophilised at −80 °C for 24 h with a freeze dryer (FreeZone 12 Plus, Labconco, MO, USA) to remove the remaining water.
5
Phenolic and Antioxidant Comparison in Popcorn
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Samples were weighed and ground to a fine powder in a SPEX Sample Prep 6750 Freezer/Mill. Samples were then transferred to a glass vial and freeze dried in a Labconco FreeZone 12 Plus freeze drier, with the collector temperature set to −80 °C, overnight to remove any water present in the sample and that might have been absorbed during the grinding process. On average samples lost less than 10% of weight after the freeze-drying period. Samples were stored at −20 °C until they were further processed.
To study the difference in phenolic content and antioxidant capacity between the pericarp and the endosperm + germ, the pericarp of raw kernels were separated with a scalpel from the rest of the kernel. The pericarp and endosperm + germ were ground as described above.
To study the effect of popping, three popcorn kernel brands were popped in a household microwave oven. The popped kernels were ground into a fine powder and processed in the same manner as whole kernels.
To study the difference in phenolic content and antioxidant capacity between the pericarp and the endosperm + germ, the pericarp of raw kernels were separated with a scalpel from the rest of the kernel. The pericarp and endosperm + germ were ground as described above.
To study the effect of popping, three popcorn kernel brands were popped in a household microwave oven. The popped kernels were ground into a fine powder and processed in the same manner as whole kernels.
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