"Per cell" normalization method for mRNA measurement by quantitative PCR and microarrays

BACKGROUND: Transcriptome data from quantitative PCR (Q-PCR) and DNA microarrays are typically obtained from a fixed amount of RNA collected per sample. Therefore, variations in tissue cellularity and RNA yield across samples in an experimental series compromise accurate determination of the absolute level of each mRNA species per cell in any sample. Since mRNAs are copied from genomic DNA, the simplest way to express mRNA level would be as copy number per template DNA, or more practically, as copy number per cell. RESULTS: Here we report a method (designated the "Percellome" method) for normalizing the expression of mRNA values in biological samples. It provides a "per cell" readout in mRNA copy number and is applicable to both quantitative PCR (Q-PCR) and DNA microarray studies. The genomic DNA content of each sample homogenate was measured from a small aliquot to derive the number of cells in the sample. A cocktail of five external spike RNAs admixed in a dose-graded manner (dose-graded spike cocktail; GSC) was prepared and added to each homogenate in proportion to its DNA content. In this way, the spike mRNAs represented absolute copy numbers per cell in the sample. The signals from the five spike mRNAs were used as a dose-response standard curve for each sample, enabling us to convert all the signals measured to copy numbers per cell in an expression profile-independent manner. A series of samples was measured by Q-PCR and Affymetrix GeneChip microarrays using this Percellome method, and the results showed up to 90 % concordance. CONCLUSION: Percellome data can be compared directly among samples and among different studies, and between different platforms, without further normalization. Therefore, "percellome" normalization can serve as a standard method for exchanging and comparing data across different platforms and among different laboratories

ミズ ヲ リヨウシタ ウチュウセン チュウセイシ カンソク ソウチ

当初,南極観測用に製作された宇宙線中性子観測装置は,IGY用として設計されたもので,全体の重量が6トンを超えるため,ヘリコプター輸送に適さない.そこで重量軽減化の方法として,(1)ニュートロン減速剤として,パラフィンの代りに現地の海水を利用する.(2)真空管方式の増幅,計数装流をすべてトランジスター化する.の2点から,新しい観測装置を製作した.この装置は重量約500kgで,第4次観測隊により無事基地へ空輪された.観測は1960年3月3日より開始された.観測結果の一部として,日平均値の変化及び5月4日の宇宙線異常増加の結果を示した

電力監視システム導入による八戸工業大学における電力の「見える化」

表紙に表示のISSNはタイトル変更に伴い、21866015に変更 第8巻までのタイトル「八戸工業大学異分野融合科学研究所紀要

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代表的なメソポーラスシリカとして知られるTi-HMS, MCM-41そしてFSM-16を有機合成反応における反応促進剤として活用することを試みた。ルイス酸サイトを有するTi-HMSが接触還元条件下,選択的に水酸基の脱ベンジル化反応を加速することを見出した。 MCM-41が水酸基の保護基として知られるt-butyldimethylsilyl (TBS)基存在下,triethylsilyl (TES)基を選択的に脱保護できることを見出した。 FSM-16がα-ハイドロキシカルボン酸の酸化的光脱炭酸反応やアリールメチルハライドやアリールメチルアルコールの光酸化反応,スチレン類の酸化開裂反応等幾つかの光反応を促進することを見出した。Ti-HMS, MCM-41 and FSM-16 which are typical hexagonal mesoporous silica, were investigated for use as a promoter for organic synthesis. Ti-HMS, which possesses Lewis acid sites, was found to accelerate deprotection of the benzyl group, which is a typical protecting group of the hydroxyl group, selectively under hydrogenolytic conditions. MCM-41 proved to deprotect the triethylsilyl (TES) group, which is a typical protecting group of hydroxyl group, selectively and easily in the presence of t-butyldimethylsilyl (TBS) group. FSM-16 was found to accelerate several photo-reactions such as oxidative photodecarboxylation of α-hydroxy carboxylic acid, oxidation of arylmethylhalides and arylmethylalcohols, and oxidative cleavage of styrenes