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NuncTM 15mL & 50mL Conical Sterile Polypropylene Centrifuge Tubes

Nunc TM 15mL&50mL圆锥形无菌聚丙烯离心管

公司名称: Thermo Fisher Scientific
产品编号: 339653
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Generation of Mutant Pigs by Direct Pronuclear Microinjection of CRISPR/Cas9 Plasmid Vectors
Author:
Date:
2017-06-05
[Abstract]  A set of Cas9 and single guide CRISPR RNA expression vectors was constructed. Only a very simple procedure was needed to prepare specific single-guide RNA expression vectors with high target accuracy. Since the de novo zygotic transcription had been detected in mouse embryo at the 1-cell stage, the plasmid DNA vectors encoding Cas9 and GGTA1 gene specific single-guide RNAs were micro-injected into zygotic pronuclei to confirm such phenomenon in 1-cell pig embryo. Our results demonstrated that mutations caused by these CRISPR/Cas9 plasmids occurred before and at the 2-cell stage of ... [摘要]  构建了一套Cas9和单引导CRISPR RNA表达载体。 需要一个非常简单的程序来制备具有高目标精度的特异性单向RNA表达载体。 由于在1细胞期的小鼠胚胎中已经检测到了合并的合子转录,所以将编码Cas9和GGTA1基因特异性单向RNA的质粒DNA载体微注射到合子原核中以确认 1细胞猪胚胎现象。 我们的研究结果表明,这些CRISPR / Cas9质粒引起的突变发生在猪胚胎的2细胞阶段之前和之后,表明除了体外转录的RNA的细胞质显微注射外,CRISPR / Cas9 DNA载体为产生基因敲除猪提供了有效的解决方案。

背景 由于最初发现了大肠杆菌基因组(Ishino)中的大肠杆菌基因组下游的32bp间隔的串联重复的高度保守的29个碱基对(bp)序列,等等,1987; Nakata等人,1989),在约50%至50bp的大小变化的短定期间隔重复序列的家族中发现约50%细菌和90%的古细菌(Makarova等人,2015)。根据它们的特征结构,Mojica(Mojica等人,2009)和Jansen(Jansen等人)引入了定期交织的短回文重复序列(CRISPR)的名称, ,2002),目前普遍使用。首先通过核苷酸序列比对(Jansen等人,2002)在CRISPR基因座的侧翼首先鉴定了一组CRISPR相关基因 cas1 ...

Nuclei Isolation from Nematode Ascaris
Author:
Date:
2017-05-05
[Abstract]  Preparing nuclei is necessary in a variety of experimental paradigms to study nuclear processes. In this protocol, we describe a method for rapid preparation of large number of relatively pure nuclei from Ascaris embryos or tissues that are ready to be used for further experiments such as chromatin isolation and ChIP-seq, nuclear RNA analyses, or preparation of nuclear extracts (Kang et al., 2016; Wang et al., 2016). [摘要]  在各种实验范例中准备核是必要的,以研究核过程。在本协议中,我们描述了一种从准备用于进一步实验(如染色质分离和ChIP-seq,核RNA)的蛔虫胚胎或组织快速制备大量相对纯的核的方法分析或准备核提取物(Kang等人,2016; Wang等人,2016)。

背景 核分离通常是研究核事件的分子和生物化学方面的第一步。已经开发了几种方法来分离来自不同组织和细胞类型的细胞核。然而,除了C以外的线虫的细胞核分离方案很少。已经描述了线索,(Ooi等人,2010; Zanin等人,2011; Haenni等人,2012, )。已经使用寄生线虫蛔虫的胚胎来制备用于体外无细胞系统的各种提取物(Cohen等人,2004) ; Lall等人,2004)。然而,这些提取物通常是全细胞提取物。在这里,我们描述了从线虫蛔虫制备细胞核的方法。

Differentiation of Human Embryonic Stem Cells into Cone Photoreceptors
Author:
Date:
2016-07-20
[Abstract]  Photoreceptors are specialized retinal neurons able to respond to light in order to generate visual information. Among photoreceptors, cones are involved in colors discrimination and high-resolution central vision and are selectively depleted in macular degenerations and cone dystrophies. A possible therapeutic solution for these disorders is to replace degenerating cells with functional cones. Here, we describe a simple protocol for the rapid production of large amount of cone photoreceptors from human pluripotent stem cells. The differentiation protocol is based on the “default pathway” of ... [摘要]  光感受器是能够响应光以产生视觉信息的专门的视网膜神经元。 在光感受器中,视锥细胞参与颜色辨别和高分辨率中心视觉,并且选择性缺失黄斑变性和视锥营养不良。 这些疾病的可能的治疗溶液是用功能性锥体代替变性细胞。 在这里,我们描述了一个简单的协议,从人类多能干细胞快速生产大量的锥形感光细胞。 分化方案基于使用BMP,TGFβ和WNT拮抗剂COCO的神经诱导的"默认途径"。

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