Name: the mplex protocol for multi-omic analyses of soil samples
Uploaded: 2018-05-30T14:00:00
Description: Watch this Scientific Journal Video about The MPLEx Protocol for Multi-omic Analyses of Soil Samples at JoVE.com

著者らは、彼について数字を準備ネイサン ジョンソンを感謝したいです。この研究は、米国エネルギー省のオフィスの生物学・環境研究 (ゲノム科学プログラム)、移行 (ミント) 研究室研究開発の内細菌叢解析によって資金を供給されるパン オミックス プログラムによって支えられました。イニシアチブは、国家機関の健康国立環境衛生研究所 (R01 ES022190) と同様、太平洋の北西国立研究所で、NIH (P42 ES027704)。KEBJ は、R21 HD084788 新規マルチ弔抽出技術開発する金融支援を感謝したいです。この作品で、w. r. ワイリー環境分子科学研究所 (EMSL)、DOE 国立科学ユーザー施設で太平洋の北西国立研究所 (なり) を行った。なり、マルチ プログラム国立研究所契約 DE AC06 76RL01830 下 DOE のバテルが運営。

注: 非常にウェット土壌は抽出の有効性を損なうことがなく抽出前に凍結乾燥することができます。湿った土も使用できますが、特定の比率で試薬を追加する場合に考慮すべき。
注: 2 つの 50 mL チューブ (最大 50 mL チューブあたり 10 g 土) 間で分割する必要があります抽出あたりの乾燥した土壌重量の 20 g を使用することをお勧めします。抽出は、使用可能なサンプルに?...

<ol>
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L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+fungus+gardens+of+leaf-cutter+ants+undergo+a+distinct+physiological+transition+during+biomass+degradation.">The fungus gardens of leaf-cutter ants undergo a distinct physiological transition during biomass degradation.</a> Environ Microbiol Rep. 6, 389-395 (2014).</li><li>Deatherage Kaiser, B. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Multi-Omic+View+of+Host-Pathogen-Commensal+Interplay+in+Salmonella-Mediated+Intestinal+Infection.">A Multi-Omic View of Host-Pathogen-Commensal Interplay in Salmonella-Mediated Intestinal Infection.</a> PLoS One. 8, e67155 (2013).</li><li>Kim, Y. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salmonella+modulates+metabolism+during+growth+under+conditions+that+induce+expression+of+virulence+genes.">Salmonella modulates metabolism during growth under conditions that induce expression of virulence genes.</a> Mol Biosyst. 9, 1522-1534 (2013).</li><li>Ansong, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+multi-omic+systems+approach+to+elucidating+Yersinia+virulence+mechanisms.">A multi-omic systems approach to elucidating Yersinia virulence mechanisms.</a> Mol Biosyst. 9, 44-54 (2013).</li><li>Bordbar, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Model-driven+multi-omic+data+analysis+elucidates+metabolic+immunomodulators+of+macrophage+activation.">Model-driven multi-omic data analysis elucidates metabolic immunomodulators of macrophage activation.</a> Mol Syst Biol. 8, 558 (2012).</li><li>Hu, Z. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metabolomic+response+of+human+skin+tissue+to+low+dose+ionizing+radiation.">Metabolomic response of human skin tissue to low dose ionizing radiation.</a> Mol Biosyst. 8, 1979-1986 (2012).</li><li>Perera, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dengue+virus+infection+perturbs+lipid+homeostasis+in+infected+mosquito+cells.">Dengue virus infection perturbs lipid homeostasis in infected mosquito cells.</a> PLoS Pathog. 8, e1002584 (2012).</li><li>Gao, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+reversed-phase+capillary+ultra-performance+liquid+chromatography-mass+spectrometry+(UPLC-MS)+method+for+comprehensive+top-down/bottom-up+lipid+profiling.">A reversed-phase capillary ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) method for comprehensive top-down/bottom-up lipid profiling.</a> Anal Bioanal Chem. 402, 2923-2933 (2012).</li><li>Sorensen, C. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perturbations+in+the+lipid+profile+of+individuals+with+newly+diagnosed+type+1+diabetes+mellitus:+Lipidomics+analysis+of+a+Diabetes+Antibody+Standardization+Program+sample+subset.">Perturbations in the lipid profile of individuals with newly diagnosed type 1 diabetes mellitus: Lipidomics analysis of a Diabetes Antibody Standardization Program sample subset.</a> Clin Biochem. 43, 948-956 (2010).</li><li>Diamond, D. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temporal+proteome+and+lipidome+profiles+reveal+hepatitis+C+virus-associated+reprogramming+of+hepatocellular+metabolism+and+bioenergetics.">Temporal proteome and lipidome profiles reveal hepatitis C virus-associated reprogramming of hepatocellular metabolism and bioenergetics.</a> PLoS Pathog. 6, e1000719 (2010).</li><li>Alquier, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deletion+of+GPR40+impairs+glucose-induced+insulin+secretion+in+vivo+in+mice+without+affecting+intracellular+fuel+metabolism+in+islets.">Deletion of GPR40 impairs glucose-induced insulin secretion in vivo in mice without affecting intracellular fuel metabolism in islets.</a> Diabetes. 58, 2607-2615 (2009).</li><li>Ding, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+the+accurate+mass+and+time+tag+approach+in+studies+of+the+human+blood+lipidome.">Application of the accurate mass and time tag approach in studies of the human blood lipidome.</a> J Chromatogr B Analyt Technol Biomed Life Sci. 871, 243-252 (2008).</li><li>Rasmussen, A. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Systems+virology+identifies+a+mitochondrial+fatty+acid+oxidation+enzyme,+dodecenoyl+coenzyme+A+delta+isomerase,+required+for+hepatitis+C+virus+replication+and+likely+pathogenesis.">Systems virology identifies a mitochondrial fatty acid oxidation enzyme, dodecenoyl coenzyme A delta isomerase, required for hepatitis C virus replication and likely pathogenesis.</a> J Virol. 85, 11646-11654 (2011).</li><li>Manza, L. L., Stamer, S. L., Ham, A. J., Codreanu, S. G., Liebler, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sample+preparation+and+digestion+for+proteomic+analyses+using+spin+filters.">Sample preparation and digestion for proteomic analyses using spin filters.</a> Proteomics. 5, 1742-1745 (2005).</li><li>Wisniewski, J. R., Zougman, A., Nagaraj, N., Mann, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Universal+sample+preparation+method+for+proteome+analysis.">Universal sample preparation method for proteome analysis.</a> Nat Methods. 6, 359-362 (2009).</li><li>Zhou, J. Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simple+sodium+dodecyl+sulfate-assisted+sample+preparation+method+for+LC-MS-based+proteomics+applications.">Simple sodium dodecyl sulfate-assisted sample preparation method for LC-MS-based proteomics applications.</a> Anal Chem. 84, 2862-2867 (2012).</li><li>Anderson, J. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+abundance+of+type+III+secretion+system-inducing+signals+in+Arabidopsis+mkp1+enhances+resistance+against+Pseudomonas+syringae.">Decreased abundance of type III secretion system-inducing signals in Arabidopsis mkp1 enhances resistance against Pseudomonas syringae.</a> Proc Natl Acad Sci U S A. 111, 6846-6851 (2014).</li><li>Chourey, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+cellular+lysis/protein+extraction+protocol+for+soil+metaproteomics.">Direct cellular lysis/protein extraction protocol for soil metaproteomics.</a> J Proteome Res. 9, 6615-6622 (2010).</li><li>Kim, S., Gupta, N., Pevzner, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectral+probabilities+and+generating+functions+of+tandem+mass+spectra:+a+strike+against+decoy+databases.">Spectral probabilities and generating functions of tandem mass spectra: a strike against decoy databases.</a> J Proteome Res. 7, 3354-3363 (2008).</li><li>Kim, S., Pevzner, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MS-GF++makes+progress+towards+a+universal+database+search+tool+for+proteomics.">MS-GF+ makes progress towards a universal database search tool for proteomics.</a> Nat Commun. 5, 5277 (2014).</li><li>Cole, J. 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それは、すべてではない所が同じ利用可能な機器特定のメソッドは、たとえば散のステップを合わせることができるので注意してください。ここで我々 はボルテックスと sonicating、ただし 50 mL の大きいビード ビーターの使用動作を使用します。-105 ° C のコレクターの温度とエアカーテンを使用できない場合は、窒素気流下におけるサンプルを乾かすことができます。また土壌の種類が大き?...

炭素循環と気候変動を理解するための重要な含意がある土壌微生物の評価最近の研究はしかし各種土壌中の細菌のゲノムをシーケンスの欠如など検出される蛋白質の多くの未知関数の難しさを強調しています。これらの課題は、1,2,3これまで知られている最も複雑な微生物群集をされている土壌から起因します。メタゲノム、metatranscriptomic、metaproteomic、メタボローム、リピドミクス研究からの結果を組み合わせると、マルチ弔分析は存在中の微生物により深い理解を得るために多数の土壌調査で最近実装されています。環境変動1,4、5のため起きている分子変化について包括的な情報を取得します。マルチ本腰を入れて研究課題の 1 つは質量分析法 (MS)-ベース metaproteomic、メタボローム、リピドミクスの測定通常各 MS の互換性6,7に本腰を入れての特定抽出処理が必要,8,9. これらの正確な手順を作るその実装非常に困難または不可能なときだけのサンプルの数量限定。これらの課題は、方法の検討を同時代謝・蛋白質・脂質抽出 (MPLEx) 小さいサンプル ボリュームまたは固まりを使用して、精度、およびすべての 3 つの解析の高速サンプル準備を提供することができる私たちを求めています。10. までに、これらの目標をすべて達成することができます代替土壌抽出手順がないです。
グローバル マルチ弔単一の土壌サンプルの分析を有効にするには、クロロホルム、メタノール、水分離に基づく有機溶媒抽出プロトコルは利用10だったこのメソッドは総脂質抽出9,11のために開発、最近では単一のサンプル12,13 代謝、蛋白質および脂質の同時抽出のため改正されました。、14,15,16,17,18,19,20,21,22、 23,24,25,26,27,28,29,30より少ないサンプル量を有効にして実験的変動10。MPLEx プロトコルでクロロホルムは異なる画分にサンプル成分の三相の化学分離のための基礎を提供する水と混和性ではありません。トップの水相にはしたがって蛋白質ディスクおよび脂質層の順下クロロホルム段階 (図 1) で親水性の代謝物が含まれます。ほとんどの土に MPLEx を適用すると、微粒子の残骸はサンプリング チューブの一番下で蓄積し、すべてのレイヤーが収集された後に破棄することができます。各土壌の種類が異なる、ただし、泥炭などの高有機質土、土の破片中間層にとどまり、サンプリング チューブの底にも該当しません。MPLEx マルチ弔解析で同じサンプル低下 2) マルチ弔抽出 1) サンプルの少量を使用できるよう、同じサンプルから型複数分子を分離するときにいくつかの利点を提供します全体的に実験的変動と3) より多くのサンプルは、高スループット研究10はるかに速く準備できます。一緒にこれらの利点が評価土壌試料および彼らの複雑な微生物群集のより優れた計測機能を提供するために不可欠です。

<table>
	<tbody>
		<tr>
			<td>Chloroform</td>
			<td>Sigma-Aldrich</td>
			<td>650498</td>
			<td>Stored at -20&#176;C !Caution chloroform has acute potential health effects, skin irritation and possible chemical burns, irritation to the respiratory system, may affect the kidneys, liver, heart. Wear suitable protective glasses, clothing and gloves, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Methanol</td>
			<td>Sigma-Aldrich</td>
			<td>34860</td>
			<td>Stored at -20&#176;C !Caution Methanol may cause respiratory tract, skin and eye irritation, may damage the nerves, kidneys and liver. Wear suitable protective glasses, clothing and gloves, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Purified water from Millipore</td>
			<td>Milli-Q</td>
			<td></td>
			<td>Water purification system.</td>
		</tr>
		<tr>
			<td>Sodium dodecyl sulfate</td>
			<td>Sigma-Aldrich</td>
			<td>L6026</td>
			<td>!Caution SDS causes acute toxicity and is flammable. It is a skin, eye and airway irritant. Wear gloves and safety glasses.</td>
		</tr>
		<tr>
			<td>Soil protein extraction kit</td>
			<td>MoBio, NoviPure Soil Protein Extraction Kit, Qiagen</td>
			<td>30000-20</td>
			<td></td>
		</tr>
		<tr>
			<td>DL-dithiothreitol</td>
			<td>Sigma-Aldrich</td>
			<td>43815</td>
			<td></td>
		</tr>
		<tr>
			<td>1M Trizma HCL</td>
			<td>Sigma-Aldrich</td>
			<td>T2694</td>
			<td></td>
		</tr>
		<tr>
			<td>Trichloroacetic acid</td>
			<td>Sigma-Aldrich</td>
			<td>T0699</td>
			<td>!Caution TCA is caustic, toxic and may cause skin burns. Wear gloves and safety glasses.</td>
		</tr>
		<tr>
			<td>Acetone</td>
			<td>Sigma-Aldrich</td>
			<td>650501</td>
			<td>Stored at -20&#176;C !Caution Acetone may cause respiratory tract and skin and eye irritation. Flammable liquid and vapor. Wear safety glasses gloves and a lab coat, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Urea</td>
			<td>Sigma-Aldrich</td>
			<td>208884</td>
			<td>!Caution Urea is an eye and skin irritant, use gloves and safety glasses</td>
		</tr>
		<tr>
			<td>Ammonium bicarbonate</td>
			<td>Fluka</td>
			<td>09830</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypsin</td>
			<td>Promega</td>
			<td>V528A</td>
			<td>20&#181;g vials</td>
		</tr>
		<tr>
			<td>Bicinchoninic acid protein assay kit</td>
			<td>Pierce</td>
			<td>23227</td>
			<td></td>
		</tr>
		<tr>
			<td>Ammonium Formate</td>
			<td>Sigma-Aldrich</td>
			<td>09735</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetonitrile</td>
			<td>Sigma-Aldrich</td>
			<td>34998</td>
			<td>!Caution Acetonitrile is a skin and eye irritant. Highly flammable. Wear gloves and safety glasses. Work in a fume hood.</td>
		</tr>
		<tr>
			<td>Trifluoroacetic acid</td>
			<td>Sigma-Aldrich</td>
			<td>T6508</td>
			<td>!Caution TFA is extremely hazardous in case of skin contact, eye contact, ingestion and inhalation. May produce tissue damage particularly on mucous membranes of eyes, mouth and respiratory tract. Skin contact may produce burns. Wear gloves, lab coat, safety glasses and work in a fume hood.</td>
		</tr>
		<tr>
			<td>Methoxyamine hydrochloride</td>
			<td>Sigma-Aldrich</td>
			<td>226904</td>
			<td>!Caution Methoxyamine hydrochloride causes severe burns and serious damage to eyes, may cause sensitization by skin contact. Wear safety glasses, gloves and lab coat, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Pyridine</td>
			<td>Sigma-Aldrich</td>
			<td>270970</td>
			<td>!Caution Pyridine can cause skin and eye irritation, central nervous system depression. Vapor may cause flash fire. Wear safety glasses, gloves and lab coat, work in a fume hood.</td>
		</tr>
		<tr>
			<td>N-Methyl-N-(trimethylsilyl)trifluoroacetamide with 1% trimethylchlorosilane</td>
			<td>Sigma-Aldrich</td>
			<td>69478</td>
			<td>!Caution MSTFA + 1% TMCS can cause skin corrosion, serious eye damage and specific target organ toxicity. Flammable liquid and vapor. Wear safety glasses, gloves and lab coat, work in a fume hood.</td>
		</tr>
		<tr>
			<td>Potassium chloride</td>
			<td>Sigma-Aldrich</td>
			<td>P9541</td>
			<td></td>
		</tr>
		<tr>
			<td>Milli-Q water purification system</td>
			<td>Millipore</td>
			<td></td>
			<td>model MPGP04001</td>
		</tr>
		<tr>
			<td>Vortex</td>
			<td>Scientific Industries</td>
			<td>SI-0236</td>
			<td>Vortex Genie 2</td>
		</tr>
		<tr>
			<td>Probe sonicator</td>
			<td>FisherBrand</td>
			<td></td>
			<td>model FB505</td>
		</tr>
		<tr>
			<td>Refrigerated centrifuge</td>
			<td>Eppendorf</td>
			<td></td>
			<td>model 5810R</td>
		</tr>
		<tr>
			<td>50mL tube swinging bucket rotor</td>
			<td>Eppendorf</td>
			<td>A-4-44</td>
			<td></td>
		</tr>
		<tr>
			<td>50mL fixed angle rotor</td>
			<td>Eppendorf</td>
			<td>FA-45-6-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Balance</td>
			<td>OHAUS</td>
			<td></td>
			<td>model V22PWE150IT</td>
		</tr>
		<tr>
			<td>Serological pipette controller</td>
			<td>Eppendorf</td>
			<td>12-654-100</td>
			<td></td>
		</tr>
		<tr>
			<td>10mL, 25mL glass serological pipettes</td>
			<td>FisherBrand</td>
			<td>13-678-27F, 13-678-36D</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermomixer with Thermotop</td>
			<td>Eppendorf</td>
			<td>5382000015, 5308000003</td>
			<td></td>
		</tr>
		<tr>
			<td>0.9 &#8211; 2.0 mm blend stainless steel beads</td>
			<td>NextAdvance</td>
			<td>SSB14B</td>
			<td></td>
		</tr>
		<tr>
			<td>0.15 mm garnet beads</td>
			<td>MoBio</td>
			<td>13122-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnetic stir plate</td>
			<td>FisherBrand</td>
			<td>11-100-16SH</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnetic stir bar</td>
			<td>FisherBrand</td>
			<td>14512130</td>
			<td></td>
		</tr>
		<tr>
			<td>pH paper strips, pH range 0&#8211;14</td>
			<td>FisherBrand</td>
			<td>M95903</td>
			<td></td>
		</tr>
		<tr>
			<td>15mL, 50mL conical polypropylene centrifuge tube</td>
			<td>Genesee Scientific</td>
			<td>21-103 21-108</td>
			<td>chloroform compatible</td>
		</tr>
		<tr>
			<td>50mL vortex attachment</td>
			<td>MoBio</td>
			<td>13000-V1-50</td>
			<td></td>
		</tr>
		<tr>
			<td>Ice bucket</td>
			<td>FisherBrand</td>
			<td>02-591-44</td>
			<td></td>
		</tr>
		<tr>
			<td>27.25x70mm glass vials</td>
			<td>FisherBrand</td>
			<td>03-339-22K</td>
			<td></td>
		</tr>
		<tr>
			<td>Breathe Easier plate membranes</td>
			<td>Midwest Scientific</td>
			<td>BERM-2000</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol wipes</td>
			<td>Diversified Biotech</td>
			<td>BPWP-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Heater shaker incubator</td>
			<td>Benchmark, Incu-Shaker Mini</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Analog rotisserie tube rotator</td>
			<td>SoCal BioMed, LLC</td>
			<td>82422001</td>
			<td></td>
		</tr>
		<tr>
			<td>Filter-Aided-Sample-Prep kit</td>
			<td>FASP; Expedeon</td>
			<td>44250</td>
			<td></td>
		</tr>
		<tr>
			<td>Microplate reader</td>
			<td>Biotek, EPOCH</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>-20 Degree Celsius Freezer</td>
			<td>Fisher</td>
			<td>13986149</td>
			<td></td>
		</tr>
		<tr>
			<td>-80 Degree Celsius Freezer</td>
			<td>Stirling Ultracold</td>
			<td>SU78OUE</td>
			<td></td>
		</tr>
		<tr>
			<td>Q-Exactive ion trap mass spectrometer</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Agilent 7890A gas chromatograph coupled with a single quadrupole 5975C mass spectrometer</td>
			<td>Agilent Technologies, Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>LTQ-Orbitrap Velo</td>
			<td>Thermo Scientific</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Waters NanoEquityTM UPLC system</td>
			<td>Millford, MA</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>250mL media bottle</td>
			<td>FisherBrand</td>
			<td>1395-250</td>
			<td></td>
		</tr>
		<tr>
			<td>Waters vial</td>
			<td>Waters</td>
			<td>186002805</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass MS sample vial and inserts</td>
			<td>MicroSolv</td>
			<td>9502S-WCV, 9502S-02ND</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass HPLC vial and snap caps</td>
			<td>MicroSolv</td>
			<td>9512C-0DCV, 9502C-10C-B</td>
			<td></td>
		</tr>
		<tr>
			<td>HPLC 96-well plate</td>
			<td>Agilent</td>
			<td>5042-6454</td>
			<td></td>
		</tr>
		<tr>
			<td>Large glass vial 27.25x70mm</td>
			<td>FisherBrand</td>
			<td>03-339-22K</td>
			<td></td>
		</tr>
		<tr>
			<td>Lyophilizer</td>
			<td>Labconco</td>
			<td>7934021</td>
			<td></td>
		</tr>
		<tr>
			<td>Polished stainless steel flat head spatula</td>
			<td>Spoonula; FisherBrand</td>
			<td>14-375-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Kim wipes</td>
			<td>Kimberly-Clark</td>
			<td>34721</td>
			<td></td>
		</tr>
		<tr>
			<td>XBridge C18, 250x4.6 mm, 5 &#956;M with 4.6x20 mm guard column</td>
			<td>Waters</td>
			<td>186003117, 186003064</td>
			<td></td>
		</tr>
		<tr>
			<td>Agilent 1100 series HPLC system</td>
			<td>Agilent Technologies</td>
			<td>G1380-90000</td>
			<td></td>
		</tr>
		<tr>
			<td>1.7mL centrifuge tube</td>
			<td>Sorenson</td>
			<td>11700</td>
			<td></td>
		</tr>
		<tr>
			<td>Hamilton Glass Syringes, 5mL, 50&#181;L and 250&#181;L</td>
			<td>Hamilton</td>
			<td>81517, 80975, 81175</td>
			<td></td>
		</tr>
		<tr>
			<td>Pasteur Pipettes</td>
			<td>FisherBrand</td>
			<td>13-678-20A</td>
			<td></td>
		</tr>
		<tr>
			<td>Pasteur Pipette Bulbs</td>
			<td>Sigma-Aldrich</td>
			<td>Z111597</td>
			<td></td>
		</tr>
		<tr>
			<td>Bath Sonicator</td>
			<td>Branson 1800 Ultrasonic Cleaner</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Vacuum Centrifuge</td>
			<td colspan="2">Labconco Centrivap Acid-Resistant Concentrator System</td>
			<td></td>
		</tr>
		<tr>
			<td>MicroSpin Columns, C18 Silica</td>
			<td>The Nest Group</td>
			<td>SEM SS18V</td>
			<td></td>
		</tr>
	</tbody>
</table>

the mplex protocol for multi-omic analyses of soil samples

質量分析法 (MS)-ベースの統合された metaproteomic、メタボローム、リピドミクス (マルチ弔) 研究私たちの能力を理解し、環境および生物学的システムにおける微生物群集の特性を変形させています。これらの測定は、最も複雑な土壌微生物の有効な解析結果を有効にしても複雑な微生物システムの日付に知られています。別々 に抽出は通常準備時間と必要なサンプル量を大幅増幅の本腰を入れて研究に必要なのでマルチ弔解析では、ただし、サンプル準備課題を必要は。この制限に対処するため、同じ土壌サンプルから代謝、蛋白質および脂質 (MPLEx) の同時抽出法 3-1 では、溶剤ベースのアプローチの適応によって作成されました。この MPLEx プロトコルは、複雑な土壌サンプルの数量限定に利用する場合も簡単でかつ多くのサンプルの種類、強健をあると証明しました。MPLEx メソッドには、生物的および環境の摂動に起きている変化を評価しながらそれぞれの微生物のコミュニティのメンバーのより良い理解を得るために必要な高速マルチ本腰を入れて測定も大きく有効になります。

カンザス ネイティブ草原土壌 (Mollisol 土) から分子を抽出する MPLEx プロトコルを使用すると、帳票分析結果 3376 ペプチド、105 の脂質および提供 102 代謝物 (すべての一意の識別子)。MPLEx プロトコルは脂質代謝12,13,14,15,16,17</su...

Watch this Scientific Journal Video about The MPLEx Protocol for Multi-omic Analyses of Soil Samples at JoVE.com

The MPLEx Protocol for Multi-omic Analyses of Soil Samples

同時に単一の土壌サンプルからの代謝、蛋白質および脂質の抽出、低下したサンプル準備時間を許して、数量限定でサンプルのマルチ弔質量分析を有効にするプロトコルが表示されます。

マルチ弔土壌試料の分析のための MPLEx プロトコル

Mass spectrometry (MS)-based integrated metaproteomic, metabolomic, and lipidomic (multi-omic) studies are transforming our ability to understand and ...

Research

JoVE Journal

Chemistry

Transport of Surface-modified Carbon Nanotubes through a Soil Column

土壌カラムを通して表面修飾カーボンナノチューブの輸送

Carbon nanotubes (CNTs) are widely manufactured nanoparticles, which are being utilized in a number of consumer products, such as sporting goods, ...

A Direct, Early Stage Guanidinylation Protocol for the Synthesis of Complex Aminoguanidine-containing Natural Products

複雑なアミノグアニジン含有天然物の合成のための直接、早期グアニジニルプロトコル

The guanidine functional group, displayed most prominently in the amino acid arginine, one of the fundamental building blocks of life, is an important ...

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

Protocol for the Synthesis of Ortho-trifluoromethoxylated Aniline Derivatives

の合成のためのプロトコル<em&gt;オルソ</em&gt; -trifluoromethoxylatedアニリン誘導体

Molecules bearing trifluoromethoxy (OCF3) group often show desired pharmacological and biological properties. However, facile synthesis of ...

A Protocol for the Production of Gliadin-cyanoacrylate Nanoparticles for Hydrophilic Coating

親水性コーティングのためのグリアジンシアノアクリレートナノ粒子の製造のためのプロトコル

This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic by a simple spray ...

A Simple and Efficient Protocol for the Catalytic Insertion Polymerization of Functional Norbornenes

機能性ノルボルネンの触媒挿入重合のためのシンプルで効率的なプロトコル

Norbornene can be polymerized by a variety of mechanisms, including insertion polymerization whereby the double bond is polymerized and the bicyclic ...

A Protocol for Safe Lithiation Reactions Using Organolithium Reagents

有機リチウム試薬を用いて安全なリチウム化反応のためのプロトコル

Organolithium reagents are powerful tools in the synthetic chemist&#39;s toolbox. However, the extreme pyrophoric nature of the most reactive reagents ...

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

使用した金ナノ粒子の効率的な放射性標識のために最適化されたプロトコル<sup&gt; 125</sup&gt; I標識アジド補綴グループ

Here, we demonstrate a detailed protocol for the radiosynthesis of a 125I-labeled azide prosthetic group and its application to the efficient ...

Facile Protocol for the Synthesis of Self-assembling Polyamine-based Peptide Amphiphiles (PPAs) and Related Biomaterials

Facile Protocol for the Synthesis of Self-assembling Polyamine-based Peptide Amphiphiles PPAs and Related Biomaterials

自己組織化ペプチドのポリアミンによる両親媒性物質 (Ppa の) と関連生体材料の合成のための安易なプロトコル

Polyamine-based Peptide Amphiphiles (PPAs) are a new class of self-assembling amphiphilic biomaterials-related to the peptide amphiphiles (PAs). ...

Automation of a Positron-emission Tomography (PET) Radiotracer Synthesis Protocol for Clinical Production

Automation of a Positron-emission Tomography PET Radiotracer Synthesis Protocol for Clinical Production

臨床用陽電子放出断層レントゲン写真撮影 (ペット) ルコース合成プロトコルの自動化

The development of new positron-emission tomography (PET) tracers is enabling researchers and clinicians to image an increasingly wide array of biological ...

Biological Samples Preparation for Speciation at Cryogenic Temperature using High-Resolution  X-Ray Absorption Spectroscopy

高分解能X線吸収分光法を用いた極低温でのスペシエーションのための生体試料調製

The study of elements with X-ray absorption spectroscopy (XAS) is of particular interest when studying the role of metals in biological systems. Sample ...