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Based on modified whole genome PCR and electrophoretic mobility shift assay , our laboratory established a new method to isolate genomic regulatory sequences using unpurified nuclear extract.

本实验室在对原有的全基因组PCR技术进行改进的基础上,结合凝胶阻滞实验,建立了筛选DNA调节片段的新方法。

The smelters purchased unpurified copper from artisanal miners – men who work on their own, often illegally, and account for a large proportion of Katanga's output – before processing it and selling it overseas.

这些冶炼厂从个体采矿者(这些人自行作业,往往是非法的,占加丹加省产出的相当大一部分)那里购买未精制铜,然后对其进行加工,并销售到海外。

The assay format based on gel-immobilization achieves a high probe density and has the ability to use unpurified PCR products for array formation.

我们初步筛选出了OLR1、MMP-1、MMP-13、MMP-11、MMP-27和MMP-L1等6个基因的7个SNPs位点用于冠心病易感基因的研究。

The test results showed that the purified Pal was 1.1 wt% and the unpurified Pal was 0.8wt% when the pressure of 7MPa, temperature of 298K.

当温度为298K,压力为7MPa时,提纯的坡缕石的储氢量达到1.1wt%,而未提纯的坡缕石储氢量为0.8wt%。

By means of the natural and unrestrained language, the free and loose structure, unconventional works constructed informal style of writing, smashed the trammels of aesthetic traditional conceptions, also displayed exciting vehemence and power, with unpurified feelings as the center of expression, unbosoming and pouring out the hearts as the lyric fundamental method.

疏放"型作品以未曾净化的情感为表现中心,以宣泄与倾诉为基本抒情手段,以自然、恣肆的语言和自由、松散的结构构建了随意的文体,在审美方式上打破了传统审美观念的束缚,显示出了激动人心的气势和力量。"

The smelters purchased unpurified copper from artisanal miners– men who work on their own,often illegally,and account for alarge proportion of Katanga's output– before processing it and selling it overseas.

这些冶炼厂从个体采矿者(这些人自行作业,往往是非法的,占加丹加省产出的相当大一部分)那里购买未精制铜,然后对其进行加工,并销售到海外。

To establish a new method to isolate genomic regulatory sequence using unpurified nuclear extract, here a new strategy was developed based on modified whole genome PCR and electrophoresis mobility shift assay. The principle is as follows. After sonication and ligation with DNA linker, human genomic DNA is amplified and labeled with isotope by whole genome PCR.

为了建立一种利用未经纯化的细胞核粗提物就能从人基因组中快速、大量筛选DNA调节片段的新方法,本研究对原有的全基因组PCR技术进行改进,结合凝胶阻滞实验,建立了筛选DNA调节片段的新方法。

The hydrogen storage capacity of purified Pal 2.3wt% which was higher than unpurified Pal 1.7wt% at the same conditions(the pressure of 7MPa, temperature of 100K).

在低温(100K)、7MPa条件下,提纯坡缕石的储氢量为2.3wt%,高于未提纯的坡缕石的储氢量(1.7wt%)。

Cell proliferation and viability were assayed 48h after transfection, and MDA-7 demonstrated selective inhibition of tumor cell growth, inhibitory rates of A549, Hela and HepG2 cell lines were 25%, 20% and 19%, respectively, but had no significant effect on human fetal kidney derived 293 cell line. Hela cell was screened by G418 for 2 weeks after pcDNA3-MDA-7 and monolayer colony was counted, its monolayer colony formation was 30% of cells transfected with pcDNA3. 0 vector. CMV-driven MDA-7 adenovirus vector was constructed. 293 showed no significant apoptosis during adenovirus packaging and the unpurified adenovirus titer was about 1×10〓pfu/ml. Cos 7, A549, Hela, HepG2 and Hep3B cell was infected with Ad-GFP at different MOI.

二。黑色素细胞分化相关蛋白-7(MDA-7)的克隆及功能研究:利用RT-PCR方法从5月龄人胚胎脾细胞扩增MDA-7的编码序列,经测序鉴定序列与文献报道一致后,与真核表达载体pcDNA3.0连接,构建pcDNA3-MDA-7表达载体,瞬时转染293、A549、Hela和HepG2细胞后抽提细胞总RNA,RT-PCR结果显示表达载体可介导MDA-7在不同细胞系中有效表达;转染48h后测定细胞增殖和活力,MDA-7可选择性抑制肿瘤细胞的增殖,对A549、Hela和HepG2细胞的抑制率分别为25%、20%和19%,但是对人胚肾来源的293细胞生长无明显影响。pcDNA3-MDA-7载体转染Hela细胞后,以G418筛选2周后计数单层细胞集落形成数,计数仅为转染pcDNA3.0空载体的细胞的30%左右。

Adenosine was used as the raw material, dissolved in triethyl phosphate and then esterified with thiophosphoryl chloride under 0~5℃ for 8h without water and oxgen, in alkaline condition provided by 2,6-dimethylpyridine, introducing thiophosphoryl ester to adenosine.Then superfluous triethylamine was added and reacted under 5℃ for 1h to synthesize rough 5'-AMPS.The unpurified product was appled to a DEAE Sephadex A-25 column and eluted with triethylammonium bicarbonate.

用腺苷为起始原料,溶解于磷酸三乙脂中后,在无水无氧环境中及2,6-二甲基吡啶提供的碱性条件下与三氯硫磷以0~5℃下发生酯化反应8h,将硫代磷酸酯引入腺苷,然后再与过量三乙胺在5℃下反应1h,合成粗产品,再用碳酸氢三乙胺作洗脱液,过DEAE Sephadex A-25凝胶柱纯化。

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