To characterize fermentation of glucose among the lab generated N6 first filial (F1) generation and the parental strains (FGSC 2223 and FGSC4825), fermentation was carried out in a 96-well format in deep-well plates. Biological replicates for each strain were collected from mycelial mats grown from spore suspension in high glucose liquid media (HGLM) with a 6 gauge punch and inoculated into 750μl of HGLM (2% glucose), sealed with aluminum ThermowellTM seals, and allowed to ferment for 7 days in 12:12 Light/Dark conditions at 25°C. All samples were performed in biological quadruplicate. After fermentation, 600 μL of media was recovered and cell debris was removed by sequential centrifugation at 13.2k rpm for 5 min. The recovered supernatant was aliquoted into HPLC vials for ethanol analysis by HPLC. HPLC quantitation was performed using a Varian ProStar HPLC with a Varian ProStar Autosampler and a Varian 356-LC Refractive Index Detector. An isocratic elution was used with an Agilent Hi-Plex H+ (300 mm x 7.7 mm ion-exchange column) with 5mM H2SO4 at a flow rate of 0.7 ml/min at 60°C. The concentration of ethanol present was determined from a standard curve based on ethanol standards with a known concentration.
Prostar autosampler
Manufactured by Agilent Technologies
The ProStar Autosampler is a laboratory instrument designed to automatically introduce samples into an analytical system, such as a chromatography system. It performs sample preparation and injection tasks with high precision and repeatability, allowing for efficient and consistent data collection.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using prostar autosampler
1
Glucose Fermentation in Fungal Strains
2
Quantification of Serum Cotinine by HPLC
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Serum was extracted and samples were stored at -20 °C until analysis. Samples were obtained prospectively and analysis was performed collectively. The standard samples (calibrators and controls) were handled similarly.
Cotinine was extracted with solid-phase extraction (SPE), and quantitation was performed by reversed-phase high-performance liquid chromatography (HPLC) with UV detection [26] [27] [28] [29] [30] [31] . Cotinine was detected with UV absorbance wavelength set at 259 nm and identified by retention time index (0.38). Internal standard used was 2-phenylimidazole. The quantitation limit of cotinine was 10 ng/ml. The intra-assay and inter-assay coefficients of variation were 8% and 13%, respectively. Cotinine test was defined as positive when cotinine concentration was ≥ 10 ng/ml. The HPLC-system consisted of Varian ternair pump, Varian ProStar auto sampler, and Varian Prostar Diode Array Detector. The columns used were Bond Elut C2-solid phase extraction columns (3 ml/200 mg) and HPLC analytical column: Inertsil C8; 3.0 × 150 mm, 5 μ. Software used for controlling the HPLC-system and data processing was Galaxie chromatography software.
The technician and the blood samplers were non-smokers. Naturally, there was an interdiction to smoke in the analytical laboratory. The results of the cotinine test were never visible for the attending doctors.
Cotinine was extracted with solid-phase extraction (SPE), and quantitation was performed by reversed-phase high-performance liquid chromatography (HPLC) with UV detection [26] [27] [28] [29] [30] [31] . Cotinine was detected with UV absorbance wavelength set at 259 nm and identified by retention time index (0.38). Internal standard used was 2-phenylimidazole. The quantitation limit of cotinine was 10 ng/ml. The intra-assay and inter-assay coefficients of variation were 8% and 13%, respectively. Cotinine test was defined as positive when cotinine concentration was ≥ 10 ng/ml. The HPLC-system consisted of Varian ternair pump, Varian ProStar auto sampler, and Varian Prostar Diode Array Detector. The columns used were Bond Elut C2-solid phase extraction columns (3 ml/200 mg) and HPLC analytical column: Inertsil C8; 3.0 × 150 mm, 5 μ. Software used for controlling the HPLC-system and data processing was Galaxie chromatography software.
The technician and the blood samplers were non-smokers. Naturally, there was an interdiction to smoke in the analytical laboratory. The results of the cotinine test were never visible for the attending doctors.
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