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dNTP Mix (10 mM each)

dNTP混合物(各10mM)

公司名称: Thermo Fisher Scientific
产品编号: R0191
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Single-step Marker Switching in Schizosaccharomyces pombe Using a Lithium Acetate Transformation Protocol
Author:
Date:
2016-12-20
[Abstract]  The ability to utilize different selectable markers for tagging or mutating multiple genes in Schizosaccharomyces pombe is hampered by the historical use of only two selectable markers, ura4+ and kanMX6; the latter conferring resistance to the antibiotic G418 (geneticin). More markers have been described recently, but introducing these into yeast cells often requires strain construction from scratch. To overcome this problem we and other groups have created transformation cassettes with flanking homologies to ura4+ and kanMX6 ... [摘要]  利用不同的选择标记来标记或突变粟酒裂殖酵母中的多个基因的能力受到仅仅两种可选择标记的历史使用的阻碍,即 +和 kanMX6 ;后者赋予抗生素G418(遗传霉素)抗性。最近已经描述了更多的标记物,但是将它们引入酵母细胞通常需要从头开始施加应变。为了克服这个问题,我们和其他团队已经创建了具有与 + 和 kanMX6 的侧翼同源性的转换盒,这使得能够有效和省时的方式在现有的突变或标记的裂变酵母菌株中交换标记。
 在这里,我们提出了裂殖酵母,粟酒裂殖酵母(Schizosaccharomyces pombe)中醋酸锌转化的单步标记转换方案。以下我们将介绍如何将 ura4 + 标记交换到kanMX6 , natMX4 或 hphMX4标记,其分别对抗生素G418,海参(clonNAT)或潮霉素B提供抗性。我们还详细介绍了如何交换营养标记的任何 MX 标记,例如 arg3 + , his3 + , leu1 + 和 ura4 + 。

背景 这种用于粟酒裂殖酵母的单步骤标记交换方案允许将标记有类型的抗生素标记的任何标记或突变的基因替换为营养标记物(含有arg3 + , + ,并且已经构建了 ura4 + )并且为任何 MX交换遗传 ura4 + ...

RNA Chromatin Immunoprecipitation (RNA-ChIP) in Caenorhabditis elegans
Author:
Date:
2014-12-20
[Abstract]  The RNA chromatin immunoprecipitation assay (RNA-ChIP) allows detection and quantification of RNA–protein interactions using in vivo cross-linking with formaldehyde followed by immunoprecipitation of the RNA–protein complexes. Here we describe the RNA–ChIP protocol that we have adapted for Caenorhabditis elegans (C. elegans) to detect interaction between the nuclear Argonaute CSR-1 (chromosome segregation and RNAi deficient) protein and its target nascent RNAs. We have used a transgenic strain expressing a recombinant long isoform of CSR-1 protein fused with ... [摘要]  RNA染色质免疫沉淀测定(RNA-ChIP)允许使用体内与甲醛交联,随后RNA-蛋白复合物的免疫沉淀来检测和定量RNA-蛋白质相互作用。 在这里我们描述我们已经适应于秀丽隐杆线虫( C. elegans )的RNA-ChIP协议以检测核Argonaute CSR-1(染色体分离和RNAi缺陷 )蛋白及其目标新生RNA。 我们使用表达与N-末端3x FLAG表位融合的CSR-1蛋白的重组长同种型的转基因菌株。

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