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T4 DNA Ligase (5 U/µL)

T4 DNA连接酶(5U /μL)

公司名称: Thermo Fisher Scientific
产品编号: EL0011
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Tethered Chromosome Conformation Capture Sequencing in Triticeae: A Valuable Tool for Genome Assembly
Author:
Date:
2018-08-05
[Abstract]  Chromosome conformation capture sequencing (Hi-C) is a powerful method to comprehensively interrogate the three-dimensional positioning of chromatin in the nucleus. The development of Hi-C can be traced back to successive increases in the resolution and throughput of chromosome conformation capture (3C) (Dekker et al., 2002). The basic workflow of 3C consists of (i) fixation of intact chromatin, usually by formaldehyde, (ii) cutting the fixed chromatin with a restriction enzyme, (iii) religation of sticky ends under diluted conditions to favor ligations between cross-linked fragments ... [摘要]  染色体构象捕获测序(Hi-C)是一种全面询问细胞核中染色质三维定位的有效方法。 Hi-C的发展可以追溯到染色体构象捕获的分辨率和通量的连续增加(3C)(Dekker et al。,2002)。 3C的基本工作流程包括(i)通常用甲醛固定完整的染色质,(ii)用限制酶切割固定的染色质,(iii)在稀释条件下重新连接粘性末端,以促进交联片段之间的连接或随机片段之间的那些和(iv)量化基因组基因座对之间的连接事件的数量(de Wit和de Laat,2012)。在最初的3C方案中,通过半定量PCR扩增对应于少量基因组位点(“一对一”)的选定连接接头来测量连接频率(Dekker et al。,2002 )。然后,染色体构象捕获芯片(4C)和染色体构象捕获碳复制(5C)技术扩展3C以分别以“一对多”或“多对多”方式计算结扎事件。 Hi-C(Lieberman-Aiden et al。,2009)最终将3C与下一代测序相结合(Metzker,2010)。此处,在再连接之前,用生物素标记的核苷酸类似物填充粘性末端以在后续步骤中富集具有连接连接的片段。然后对Hi-C文库进行高通量测序,并将得到的读数映射到参考基因组,允许以“多对多”方式确定接触概率,其分辨率仅受限制性位点的分布限制和阅读深度。 Hi-C的首次应用是阐明人类基因组中的全球染色质折叠原理(Lieberman-Aiden et ...

Detection of Pathogens and Ampicillin-resistance Genes Using Multiplex Padlock Probes
Author:
Date:
2017-08-20
[Abstract]  Diagnostic assays for pathogen identification and characterization are limited either by the number of simultaneously detectable targets, which rely on multiplexing methods, or by time constraints due to cultivation-based techniques. We recently presented a 100-plex method for human pathogen characterization to identify 75 bacterial and fungal species as well as 33 clinically relevant β-lactamases (Barišić et al., 2016). By using 16S rRNA gene sequences as barcode elements in the padlock probes, and two different fluorescence channels for species and antibiotic resistance ... [摘要]  用于病原体鉴定和表征的诊断测定法由依赖于多重方法的同时可检测目标的数量或由于基于培养的技术的时间限制来限制。 我们最近提出了一种用于人类病原体鉴定的100plex方法,以鉴定75种细菌和真菌物种以及33种临床相关β-内酰胺酶(Barišić等,2016)。 通过使用16S rRNA基因序列作为挂锁探针中的条形码元件,以及用于物种和抗生素抗性鉴定的两种不同的荧光通道,我们设法将需要的微阵列探针的数量减少一半。 因此,我们在这里介绍一个运行时间约为的测定方案。 8 h,检测限为105 cfu ml-1。 正确鉴定了89%的β-内酰胺酶和93.7%的物种。
【背景】β-内酰胺酶是一类提供抗β-内酰胺抗生素的抗生素抗性基因,其结构模拟D-丙氨酰-D-丙氨酸,细菌细胞壁的一个组分,从而抑制细菌细胞壁合成。 β-内酰胺酶能够水解β内酰胺抗生素β-内酰胺环的中心成分,并使其无效(Kong et al。,2010)。今天,描述了超过1000种β-内酰胺酶,并且存在巨大的潜在环境储层(Bush,2010; Brandt等,2017)。 ...

Plant Sequence Capture Optimised for Illumina Sequencing
Author:
Date:
2014-07-05
[Abstract]  Plant Sequence Capture is used for targeted resequencing of whole exomes (all exons of a genome) of complex genomes e.g. barley and its relatives (Mascher et al., 2013). Sequencing and computing costs are significantly reduced since only the greatly enriched and gene-coding part of the barley genome is targeted, that corresponds to only 1-2% of the entire genome. Thus, applications such as genetic diversity studies and the isolation of single genes (“cloning-by-sequencing”) are greatly facilitated. Here, a protocol is provided describing the construction of shotgun DNA ... [摘要]  植物序列捕获用于复杂基因组(例如大麦及其亲属)的整个外显子(基因组的所有外显子)的靶向重测序(Mascher等人,2013)。测序和计算成本显着降低,因为只有大麦基因组的大量富集和基因编码部分被靶向,其仅对应于整个基因组的1-2%。因此,大大促进了诸如遗传多样性研究和单个基因的分离("通过测序克隆")的应用。这里,提供了描述来自基因组大麦DNA的Shotgun DNA文库的构建以在Illumina HiSeq/MiSeq系统上测序的方案。鸟枪DNA测序文库与包含大麦整个外显子组的寡核苷酸池(Exome Library)杂交。外显子组库作为包含生物素化探针(Roche/NimbleGen)的液体阵列提供。随后,使用链霉亲和素包被的磁珠对与Exome文库杂交的基因组鸟枪DNA片段进行亲和纯化。捕获的文库被PCR扩增和测序,使用高通量短读序列合成

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