GNPS - UM Matthaei botanical garden & Herbarium (low m/z)
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Orbitrap Fusion
ORGANISM(S): Viridiplantae (ncbitaxon:33090)
SUBMITTER: 
Desnor Chigumba
PROVIDER: MSV000092820 | GNPS |
REPOSITORIES: GNPS
Similar Datasets
Project description:Plant metabolic extracts. Plant tissues as specified in file name were collected from adult live plants in the University of Michigan Matthaei Botanical Garden. 0.2g fresh weight tissue were frozen, ground in a Tissuelyzer for 2min at 6m/s with 10 2.3mm silica beads in 2mL vials. Ground tissue was extracted with 3mL methanol for 1h at 37 celsius shaking. Methanol was separated from insoluble material and dried under nitrogen. Dried crude methanol extracts were resuspended in water, partitioned twice with 3 mL hexane, 3 mL ethyl acetate and finally extracted with 3 mL n-butanol. n-butanol extracts were dried in a speedvac concentrator, resuspended in 1 mL methanol, filtered through a 0.2 micron filter and analyzed by LC-MS/MS on an Thermo QExactive orbitrap with a ESI source. LC-MS settings were as follows: Injection volume 5 uL, LC - Phenomenex Kinetex 2.6 um C18 reverse phase 100 A 150 x 3 mm LC column, LC gradient: solvent A - 0.1% formic acid, solvent B - acetonitrile (0.1% formic acid), 0 min: 10% B, 5 min: min: 60% B, 5.1 min: 95% B, 6 min: 95% B, 6.1 min: 10% B, 9.9 min: 10% B, 0.5 mL/min, MS - positive ion mode, Full MS: Resolution 70000, mass range 400-1200 m/z, dd-MS2 (data-dependent MS/MS): resolution 17500, AGC target 1e5, Loop count 5, Isolation width 1.0 m/z, Collision energy 25 eV, dynamic exclusion 0.5 s.
2021-07-23 | MSV000087872 | GNPS
Project description:Plant metabolic extracts. Plant tissues as specified in file name were collected from adult live plants in the University of Michigan Matthaei Botanical Garden. 0.2g fresh weight tissue were frozen, ground in a Tissuelyzer for 2min at 6m/s with 10 2.3mm silica beads in 2mL vials. Ground tissue was extracted with 3mL methanol for 1h at 37 celsius shaking. Methanol was separated from insoluble material and dried under nitrogen. Dried crude methanol extracts were resuspended in water, partitioned twice with 3 mL hexane, 3 mL ethyl acetate and finally extracted with 3 mL n-butanol. Ethylacetate and n-butanol extracts were dried in a speedvac concentrator, resuspended in 1 mL methanol, filtered through a 0.2 micron filter and analyzed by LC-MS/MS on an Thermo QExactive orbitrap with a ESI source. LC-MS settings were as follows: Injection volume 5 uL, LC - Phenomenex Kinetex 2.6 um C18 reverse phase 100 A 150 x 3 mm LC column, LC gradient: solvent A - 0.1% formic acid, solvent B - acetonitrile (0.1% formic acid), 0 min: 10% B, 5 min: min: 60% B, 5.1 min: 95% B, 6 min: 95% B, 6.1 min: 10% B, 9.9 min: 10% B, 0.5 mL/min, MS - positive ion mode, Full MS: Resolution 70000, mass range 400-1200 m/z, dd-MS2 (data-dependent MS/MS): resolution 17500, AGC target 1e5, Loop count 5, Isolation width 1.0 m/z, Collision energy 25 eV, dynamic exclusion 0.5 s. [doi:10.25345/C5H41JX71] [dataset license: CC0 1.0 Universal (CC0 1.0)]
| MSV000092992 | GNPS
Project description:Pretreated human follicular fluid was prepared for targeted metabolomics experiments. For this purpose, 12-point standard calibration curves in the respective range with reference material were measured, and the limit of detection (LOD) and limit of quantification (LOQ) for each neurotransmitter under investigation were calculated. LC-MS/MS was conducted by an ultra-performance liquid chromatography method using a Waters ACQUITY UPLC BEH C18 column (2.1 mm×100 mm, 1.7 µm, Waters, USA). The Waters acquisition UPLC was coupled to an API 4000 triple quadrupole mass spectrometer (AB SCIEX, USA). The mobile phase A buffer was 10% methanol in water (containing 0.1% formic acid), and the mobile phase B buffer was 50% methanol in water (containing 0.1% formic acid). The LC gradient elution was at a flow rate of 0.3 mL/min at 0-8.0 min and 0.4 ml/min at 8.0-17.5 min, then 10%-30% B at 0-6.5 min; 30%-100% B at 6.5-7 min; 100% B at 7-14min; and 100%-10% B at 14-17.5min. The injection volume was 5 µl and the column temperature was set to 40 °C. Twenty-three metabolites (Ach: Acetylcholine chloride; Gln: Glutamine; Glu: Glutamate; His: L-Histidine; NE: norepinephrine; Tyr:Tyrosine; Trp:Tryptophan; Kyn: Kynurenine; GABA: 4-Aminobutyric acid;HisA:Histamine; PA:Picolinic acid;TyrA:Tyramine;DA:Hydroxytyramine hydrochloride; TrpA:Tryptamine;5-HT:Serotonin hydrochloride;E:Adrenaline hydrochloride;KynA:Kynurenic acid;5-HIAA:5-Hydroxyindole-3-acetic acid;DOPA:Levodopa;XA:Xanthurenic acid;VWA:Vanillymandelic Acid; 5-HTTP:5-Hydroxytryptophan;MT:Melatonine)were simultaneously reported in LC-MS/MS multiple reaction monitoring (MRM) mode. Data analysis was performed with Analyst 1.6 software.
2022-08-12 | PXD031560 | Pride
Project description:All chemicals were from Fisher Scientific unless otherwise noted. All solvents for extraction were HPLC grade, all solvents for mass spectrometry analysis were LCMS (Thermo Optima) grade. Streptomyces ansochromogenes subspecies pallens NRRL B-12018 was obtained from the ARS Culture Collection of the United States Department of Agriculture. A 30 mL Streptomyces ansochromogenes subspecies pallens NRRL B-12018 was inoculated from an ISP2 agar culture and cultivated in liquid ISP2 medium (4 g yeast extract, 10 g malt extract, 4 g D-glucose in 1 L deionized water, pH 7.3) for 11 days at 30 celsius at 225 rpm. The medium was then centrifuged in 50 mL centrifuge tube at 3200 g for 30 min. The cell pellet and supernatant were separated subsequently, and the supernatant (30 mL) was extracted three times with 50 mL ethyl acetate to remove lipophilic components. Then the supernatant was dried by rotary evaporation at 40 celsius. Finally, the dried supernatant was resuspended in 3.5 mL deionized water, centrifuged for 5 min at 16000 g, filtered with a Cytiva syringeless filter (0.2 micron mesh size, Cytiva, US503NPEORG). The filtered supernatant was immediately analyzed by liquid-chromatography-tandem mass spectrometry on a Thermo QExactive orbitrap mass spectrometer with a H-ESI-II ion source coupled to a Vanquish HPLC system. The LC parameters were as follows: Phenomenex Kinetex 2.6 micrometer C18 reverse phase 100 Ang. 150 x 3 mm LC column, solvent A: 20mM ammonium formate (pH 3.2), solvent B: 90% acetonitrile and 10% 200 mM ammonium formate (pH 3.2), column compartment temperature 30 celsius, injection volume 5 microliter, flow rate 0.5 mL/min, 0 min: 0% B, 5 min: 40% B, 4.5 min: 95% B, 5.5 min: 95% B, 6 min: 0% B, 10 min: 0% B. Electrospray ion source (Thermo H-ESI-II source) parameters were as follows: negative ion mode, sheath gas flow 35 psi, aux gas flow 3 psi, electrospray capillary temperature 320 celsius, electrospray capillary voltage 3.6 kV. MS parameters were as follows: full MS, resolution 70000, mass range 400-1200 m/z, dd-MS2 (data-dependent MS/MS), resolution 17500, AGC target 1x10^5, loop count 5, isolation width 1.0 m/z, collision energy 25 eV and dynamic exclusion 0.5 s. LC-MS data were analyzed with QualBrowser in the Thermo Xcalibur software package (v.4.3.73.11, Thermo Scientific).
2024-07-17 | MSV000095359 | MassIVE
Project description:Using cell sorting technique, we collected Arabidopsis root hair protoplasts (transgenic line harboring root hair specific gene promoter ::GFP, here named GFP) and nonGFP protoplasts( whole root except root hair). Then, we extracted total proteins and RNA for proteome and RNASeq. RNAseq data have already been deposited into NCBI (accession numbers SRR364677 and SRR364678). Total proteins were separated by 1D SDS-PAGE, followed by gel slicing, in gel digested and run ESI-LC-MS/MS on LTQ Orbitrap Velos. We set two biological repeats and each sample was scanned two times (two technique repeats). We then combined all ms/ms raw data from all fractions and technique repeats from one biological sample into one file to indentify proteins by searching Arabidopsis protein database with software Proteome Discoverer (with two searching engines mascot and sequester). Protocol: Protoplasts from EXP7 and non-GFP protoplastswere stored at -80°C, thawed on ice for 5 min, vortexed several times and suspended in 5 x volume of pre-cooled acetone (-20°C) containing 10% (v/v) trichloroacetic acid (TCA) and 0.07% (v/v) 2-mercaptoethanol. Proteins were then precipitated for 2 h at −20°C after thorough mixing. Proteins were collected by centrifuging at 35,000 g (JA-20 108 rotor, Beckman J2-HS) at 4°C for 30 min. The supernatant was removed and the protein pellets were washed three times with cold acetone containing 0.07% (v/v) 2-mercaptoethanol and 1 mMphenylmethanesulfonylfluoride. Protein pellets were dried by lyophilization and stored at -80°C, or immediately extracted using Laemmli buffer (63 mM TrisHCl pH 6.8, 10% glycerol, 2% SDS, 0.0025% bromophenol blue). Protein concentration was determined using Pierce protein assay kit (Pierce, Rockford). Proteins from each sample were separated using a Bis-Tris precast gradient gel (4−16%, Bio-Rad) according to the manufacturer’s instructions. After electrophoresis, the gel was Coomassie-stained using the Colloidal Blue Staining Kit (Invitrogen). The protocol used for in-gel trypsin digestion of proteins in gels was adapted from the method described in Shevchenko et al. (1996). Briefly, protein bands were manually excised and each band was cut into small pieces (~0.5 mm). The gel pieces were washed three times with a solution containing 50% methanol and 5% acetic acid for 2 h, twice with a solution containing 25 mM NH4HCO3 in 50% acetonitrile for 10 min each, and then the gel pieces were dried in a vacuum centrifuge. Reduction of proteins in gel pieces was performed with DTT and alkylation with iodoacetamide, and the gel pieces were washed and dried in a vacuum centrifuge. A trypsin solution in 25 mM NH4HCO3 containing 75 to 100 ng of sequencing grade modified trypsin (Promega) in 25-40 µl was added and incubated with gel pieces for 12-16 h at 37°C. To recover the tryptic peptides, a solution of 30 µl containing 5% formic acid and 50% acetonitril was added to the gel pieces, agitated in a vortex for 30-60 min and withdrawn into a new tube. The recovery was repeated once with 15 µl solution and the resulting two recovers were combined and dried in a vacuum centrifuge. The dried pellet was re-dissolved in 10-20 µl of 0.1% formic acid for LC-MS/MS analysis.
2013-05-28 | PXD000195 | Pride
Project description:Flag-YBX1 overexpressed T24 cells pellets were resuspended with 2 volume of lysis buffer (150 mM KCl, 10 mM HEPES pH 7.6, 2 mM EDTA, 0.5% NP-40, 0.5 mM DTT, 1:100 protease inhibitor cocktail, 400 U/ml RNase inhibitor), and incubated at 4 °C for 30 min with rotation. Then the lysate was centrifuged at 15 000 g for 20 min. Before incubating the lysate with Flag beads, 100ul were taken as input. The anti-Flag M2 magnetic beads (Sigma, 10 μl per mg lysate) were washed with NT2 buffer (200 mM NaCl, 50 mM HEPES pH 7.6, 2 mM EDTA, 0.05% NP-40, 0.5 mM DTT, 200 U/ml RNase inhibitor) four times. Cell lysate was mixed with M2 beads and incubated at 4 °C for 4 h with rotation. The beads were washed two times with 1 ml ice-cold NT2 buffer. Then the beads were subject to Micrococal nuclease (NEB) digestion (1:1 000 000 dilution) for 8 min at 37 °C. The beads were cooled on ice immediately for 5 min and washed two times with 1 ml ice-cold 1× PNK+EGTA buffer (50 mM Tris-HCl pH 7.5, 20 mM EDTA, 0.05% NP-40, 200 U/ml RNase inhibitor) and two times with 1 ml ice-cold 1× PK buffer (50 mM NaCl, 100 mM Tris-HCl pH 7.5, 10 mM EDTA, 0.2% SDS, 200 U/ml RNase inhibitor). Then the beads were digested with 200 μl pre-heated (20 min at 50 °C) Proteinase K and RNAs were extracted with an equal volume of Acid-Phenol: Chloroform, pH 4.5 (Ambion). The RNAs were subjected to rRNA removal and Bisseq. The libraries were sequenced on the Illumina HiSeq X-Ten platform at Novogene (Tianjin, CA) with paired-end 150 bp read length.The m5C sites were called using meRanCall from meRanTK (FDR < 0.01).
2019-07-02 | GSE133625 | GEO
Project description:The plant leaves were incubated (twice) at room temperature in NaCl solution (1M, 5 mL) for 15 min each, in methanol (5 mL), then in methanol-chloroform (1:1, 5 mL) twice for 30 min, and finally washed with methanol (5 mL). After each incubation, the supernatant was discarded.
The residue was dried in the open air and submitted to alkaline hydrolysis for 18 h at room temperature and in absence of light using NaOH (1M, 50 mL g-1). The alkaline extract was filtered and acidified with HCl until pH 3 and filtered again. The extract was submitted to the same process of cleanup with SPE described above, but the column was washed with ultrapure water and methanol 5% before elution using 10 mL of methanol.
2023-11-25 | MSV000093498 | MassIVE
Project description:<p>The sample to be tested was fully ground into powder in the grinder, 50 mg of the sample was freeze-dried, 1000 μL of the extraction solution containing the inner target was added (methanol:acetonitrile:water, 2:2:1, v/v/v, internal standard concentration 20 mg/L) and vortex mixed for 30 s; then add the steel ball to the 45 Hz grinder for 10 min, ultrasonic 10 min; the sample to be tested is obtained after filtration. When detecting metabolites, metabolite determination was performed based on the LC-MS system, which mainly consists of Waters Acquity I-Class PLUS ultra-high performance liquid tandem and Waters Xevo G2-XS QTof high-resolution mass spectrometer. Meanwhile, the Waters Acquity UPLC HSS T3 column (1.8 um 2.1 x 100 mm) was used as the chromatographic column. Positive and negative ion modes were used to determine the metabolites. Mobile phase A: 0.1% formic acid aqueous solution; Mobile phase B: 0.1% acetonitrile formate. The mobile phase conditions of liquid chromatography were as follows: the flow rate was 400 μL/min, 0.0 min: 98% flow A, 2% flow B; 0.25 min: 98% flow A, 2% flow B, 10.0 min: 2% flow A, 98% flow B; 13.0 min: 2% flow A, 98% flow B; 13.1 min: 98% flow A, 2% flow B; 15.0 min: 98% flow A, 2% flow B. MSe mode controlled by acquisition software (MassLynx v4.2, Waters) was used for primary and secondary mass spectrum data acquisition. ESI ion source parameters are as follows: Capillary voltage, 2500 V (positive ion mode) or -2000 V (negative ion mode); Cone hole voltage, 30 V; Ion source temperature, 100 °C; Desolvent temperature, 500 °C; Air flow rate, 50 L/h; Desolvent gas flow rate, 800 L/h; Plastic-nucleus ratio (m/z) collection range 50-1200 m/z. In the qualitative and quantitative analysis of metabolites, the original data collected by MassLynx v4.2 were processed by the Progenesis QI software for peak extraction, peak alignment, and other data, and the metabolites were identified based on the Progenesis QI software online METLIN database. Then, based on the results of the total score, MS2 score, and mass error (ppm), the metabolites were qualitatively determined.</p>
2024-07-12 | MTBLS10002 | MetaboLights
Project description:We used complementary DNA microarray to analyze the gene expression profiles in different proglottids of Moniezia expansa. A total of 4056 sequences including full length and partial complementary DNAs representing novel, known, and control genes were analyzed. We utilized cDNA array for detection of gene expression profiles of different proglottids of M.expansa. The present study provides some interesting data to better understanding the mechanisms of procreation and may suggest some potential target molecules for a more effective treatment on verminosis. 1.Construction of microarray By clustering the 2,642 sequences from the cDNA library, 1,078 unigenes of M.expansa, including full-length and partial cDNAs representing novel or known genes, were obtained for array construction. The negative control spots of non-tapeworm origin in the chip were the rice genes (48 spots). In addition, the array included 9 known genes(54 spots) as positive control genes that were provided by our library and 24 empty spots. Unigenes and all control genes were cloned into plasmid vector. The DZH (TC) chips were constructed by United Gene Holdings Limited of China (Shanghai, China) according to the method described by Li et al. The cDNA inserts were amplified by use of the polymerase chain reaction (PCR) using universal primers to plasmid vector sequences and were then purified. All PCR products were examined by agarose gel electrophoresis to ensure the quality and the identity of the amplified clones as expected. Then the amplified PCR products were dissolved in a buffer containing 3×SSC solution. The solution with amplified PCR products were spotted onto glass slides (model DZH-TC) using a Cartesian PixSys 7500 motion control robot (Cartesian Technologies, Irvine, CA., USA) fitted with ChipMaker Micro-Spotting Technology (TeleChem International, Sunnyvale, CA., USA). The glass slides were then hydrated for 2 h in 70% humidity, dried for 0.5 h at room temperature, and UV crosslinked (65 mj/cm). They were further processed at room temperature by soaking in 0.2% sodium dodecyl sulfate (SDS) for 10 min, distilled H2O for 10 min, and 0.2% sodium borohydride (NaBH4) for 10 min. The slides were dried again and ready for use. 2.RNA preparation and probe labeling Total RNAs were isolated from 6 M.expansas with scolex-neck proglottids, immature proglottids, mature proglottids and gravid proglottids. Tissue samples were ground into a fine powder in a 10 cm ceramic mortar (RNase-free) and were then homogenized in TRIzol (Biostar, Shanghai, China). After centrifugation, the supernatant was separated from the organic phase and was extracted in an equal volume of chloroform. The aqueous phase was then precipitated by an equal volume of isopropanol at 4°C, centrifuged to pellet the RNA and dissolved in Milli-Q H2O. The fluorescent cDNA probes were prepared through reverse transcription with Cy3- or Cy5-deoxy UTP (Amersham Pharmacia Biotech, Piscataway, NJ, USA) as follows: 5 μg of oligo(dT18) was added and annealed to 3μg of mRNA by heating the mixture to 70˚C for 10 min, and then chilling it on ice. The final reaction buffer mixture contained dNTPs (200 μM dATP, dCTP and dGTP; 60 μM dTTP; and 60 μM Cy3- or Cy5-dUTP), 2 μl of Superscript II reverse transcriptase (Invitrogen, Carlsbad, CA, USA) and 1×reaction buffer 10μl. The reactions were carried out at 42 ºC for 2 h. RNA was hydrolyzed by adding 4 μl of 2.5 M NaOH, incubating the mixture at 65 ºC for 10 min and then neutralizing it with 4 μl of 2.5 M HCl. The RNA samples from the scolex-neck proglottids were labeled with Cy3-dUTP and those from immature proglottids, mature proglottids and gravid proglottids, respectively, were labeled with Cy5-dUTP. The two color probes were then mixed and diluted to 500 μl with TE, and concentrated using a Microcon YM-30 filter (Millipore, Bedford, MA, USA) to 10 μl. The sample was then vacuum dried. 3.Hybridization The probe was dissolved in 20 ml of hybridization solution (5×SSC (0.75M NaCl and 0.075M sodium citrate), 0.4% SDS, 50% formamide). Microarrays were pre-hybridized with a hybridization solution containing 0.5 mg/ml of denatured salmon sperm DNA at 42°C for 6 h. Fluorescent probe mixtures were denatured at 95°C for 5 min and then applied onto the pre-hybridized chip under a cover glass. Chips were hybridized at 42°C for 15-17 h. Next, the hybridized chips were each washed at 60°C for 10 min in solutions of 2×SSC and 0.2% SDS, 0.1×SSC and 0.2% SDS, 0.1×SSC and then dried at room temperature. 4.Detection and analysis The chips were scanned with a ScanArray 4000 (GSI Lumonics, Bellerica, MA, USA) at two wavelengths, 635 and 532 nm, to detect emission from both Cy5 and Cy3, respectively. The acquired images were analyzed using GenePix Pro 3.0 software. The intensities of each spot at the two wavelengths represent the quantity of Cy3-dUTP and Cy5-dUTP. Ratios of Cy5 to Cy3 were computed using the GenePix Pro 3.0 median of ratio method. Overall intensities were normalized using the corresponding GenePix default normalization factor. All spots flagged “Bad” or “Not Found” by GenePix software were removed from the final data. Only genes with raw intensity values for both Cy3 and Cy5 of >200 were chosen for differential analysis. Genes were identified as differentially expressed if the ratio was >2 or <0.5, or the absolute value of base 2 logarithm of the ratio was >1 or < -1.
2009-12-01 | GSE13892 | GEO
Project description:Streptomyces cinnamoneus NRRL B-24434 and Streptomyces aureoverticillatus NRRL B-3326 were cultivated in ISP2 liquid medium for 7 days at 28C. This was followed by n-butanol liquid:liquid extraction of the growth medium, rotovaporation, resuspension in 80% methanol, filtration with Whatman 0.2 micron syringeless filters followed by LC-MS/MS analysis. LC-MS data was collected on a Thermo QExactive Orbitrap connected to a Vanquish LC system. LC settings were as follows: injection volume 5 microliter, Phenomenex Kinetex 2.6 micrometer C18 reverse phase 100 A 150x3 mm LC column, LC gradient, solvent A, 0.1% formic acid, solvent B, acetonitrile (0.1% formic acid), 0 min, 10% B, 5 min, 60% B, 5.1 min, 95% B, 6 min, 95% B, 6.1 min, 10% B, 9.9 min, 10% B, 0.5 ml/min. MS settings were as follows - positive ion mode, full MS, resolution 70000, mass range 400-1200 m/z, dd-MS2 (data-dependent MS/MS), resolution 17500, AGC target 1x10^5, loop count 5, isolation width 1.0 m/z, collision energy 25 eV, dynamic exclusion 0.5 s.
2024-02-11 | MSV000094063 | GNPS