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N-Lauroylsarcosine sodium salt


Company: Sigma-Aldrich
Catalog#: L5125
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Terminal Restriction Fragments (TRF) Method to Analyze Telomere Lengths
[Abstract]  Chromosome ends - telomeres - are a focus of intensive research due to their importance for the maintenance of chromosome stability. Their shortening due to incomplete replication functions as a molecular clock counting the number of cell divisions, and ultimately results in cell-cycle arrest and cellular senescence. Determination of telomere lengths is an essential approach in telomere biology for research and diagnostic applications. Terminal Restriction Fragments (TRF) analysis is the oldest approach to analyze telomere lengths and remains the “gold standard” even in current studies. This technique relies on the fact that repeated minisatellite telomeric units do not contain target sites for restriction enzymes. Consequently, telomeres remain in relatively long fragments (TRF), whereas ... [摘要]  染色体末端 - 端粒是密集研究的焦点,因为它们对维持染色体稳定性的重要性。它们由于不完全复制而缩短作为计数细胞分裂数目的分子时钟,最终导致细胞周期停滞和细胞衰老。端粒长度的测定是端粒生物学中用于研究和诊断应用的基本方法。末端限制性片段(TRF)分析是分析端粒长度的最古老的方法,并且即使在目前的研究中仍然是"金标准"。该技术依赖于重复的小卫星端粒单元不含有限制酶的靶位点的事实。因此,端粒保持相对长的片段(TRF),而基因组DNA被消化成短片段。然后通过与放射性标记的端粒探针杂交显现端粒DNA的片段。由于TRF除了端粒外还包括直到第一限制性位点的端粒相关DNA的短区域,结果稍微偏向更高的TRF值。因此,建议使用频繁的刀具或其混合物,以尽量减少这种差异。此外,通过使用TRF分析,可以区分真正(末端)端粒与间质端粒重复(ITR)(Richards和Ausubel,1988)。在该方法中,首先将BAL31消化应用于高分子量DNA。酶从其末端逐渐降解线性DNA。然后用一种或多种限制酶消化降解的DNA,并通过凝胶电泳分离片段。印迹后,用末端标记序列或端粒序列探测膜。真正的TRF可以区别于ITR,因为它们随着BAL31消化时间的增加而逐步缩短,而ITR是BAL31抗性的。在时间零时的TRF BAL31消化模式表示近似端粒长度(Fajkus等人,2005)。

Cell Fractionation of Pseudomonas aeruginosa
[Abstract]  Pseudomonas aeruginosa is a Gram negative bacterium. Separating the cell envelope compartments enables proteins to be localized to confirm where in the cell they function. Cell fractionation can also provide a first step in a protein purification strategy (Williams et al., 1998). This protocol has been designed to obtain the different fractions of P. aeruginosa, namely the inner membrane, outer membrane, cytoplasmic and periplasmic compartments. Specific detection of the arginine specific autotransporter (AaaA) (Luckett et al., 2012) in the outer membrane of P. aeruginosa has been performed using this protocol. [摘要]  铜绿假单胞菌是革兰氏阴性细菌。 分离细胞包膜区室使得蛋白质能够被定位以确认它们在细胞中的何处起作用。 细胞分级还可以提供蛋白质纯化策略的第一步(Williams等人,1998)。 该方案已经设计成获得不同比例的P。 铜绿假单胞菌,即内膜,外膜,细胞质和周质区室。 在P的外膜中特异性检测精氨酸特异性自转运体(AaaA)(Luckett等人,2012)。 已经使用该协议进行了铜绿假单胞菌。

Neutral Comet Assay
[Abstract]  The Comet assay (or Single Cell Gel Electrophoresis assay) is a sensitive technique to detect DNA damage at the level of an individual cell. This technique is based on micro-electrophoresis of cells DNA content. Briefly, cells are embedded in agarose, lysed and submitted to an electric field, before the staining step with a fluorescent DNA binding dye. Damaged DNA (charged DNA) migrates in this field, forming the tail of a “comet”, while undamaged DNA remained in the head of the “comet”. The following document describes the protocol to realize a neutral comet assay. This assay can be applied to different cell types and has been useful for numerous applications in fields of toxicology or DNA damage and repair. [摘要]  彗星测定(或单细胞凝胶电泳测定)是在单个细胞水平检测DNA损伤的敏感技术。 这种技术是基于细胞DNA含量的微电泳。 简言之,将细胞嵌入琼脂糖中,裂解并在用荧光DNA结合染料染色步骤之前将其送入电场。 损伤的DNA(带电DNA)在该领域迁移,形成“彗星”的尾部,而未损坏的DNA保留在“彗星”的头部。 以下文件描述了实现中性彗星测定的方案。 该测定可应用于不同的细胞类型,并且可用于毒理学或DNA损伤和修复领域的许多应用。