{{'Search' | translate}}
 

Microcentrifuge

5424/5424 R 离心机

Company: Eppendorf
Catalog#: 5424
Bio-protocol()
Company-protocol()
Other protocol()

Safe DNA-extraction Protocol Suitable for Studying Tree-fungus Interactions
Author:
Date:
2020-06-05
[Abstract]  We present a safe and low-cost method suitable for DNA extraction from mycelium and tree tissue samples. After sample preparation, the extraction takes about 60 min. Method performance was tested by extracting DNA from various tree tissue samples and from mycelium grown on solid and liquid media. DNA was extracted from juvenile and mature host material (Picea abies, Populus trichocarpa, Pseudotsuga menziesii) infected with different pathogens (Heterobasidion annosum, Heterobasidion parviporum, Leptographium wagenerii, Sphaerulina musiva). Additionally, DNA was extracted from pure cultures of the pathogens and several endophytic fungi. PCR success rate was 100% for young poplar material and fungal samples, and 48-72% for ... [摘要]  [摘要 ] 我们提出了一种安全,低成本的方法,适用于从菌丝体和树木组织样品中提取DNA。样品制备后,提取过程大约需要60分钟。通过从各种树木组织样品和菌丝体中提取DNA来测试方法性能DNA从少年提取和成熟主体材料(云杉冷杉,胡杨毛果,黄杉花旗松感染不同病原体()Heterobasidion annosum ,Heterobasidion parviporum ,Leptographium wagenerii ,Sphaerulina musiva )。额外的,DNA从提取纯的病原体和几种内生真菌培养物。年轻杨树材料和真菌样品的PCR成功率是100%,针叶树和成熟阔叶植物样品的PCR成功率是48-72%。我们建议使用10-50 mg的新鲜样品最好结果。该方法为研究树-真菌相互作用提供了一种安全且低成本的DNA提取方法,并且是潜在的提纯方法。兴的目的。

[背景 ] DNA提取是植物-微生物相互作用研究和植物疾病诊断purposes.The丰植物真菌相互作用的中央技术由高数量从植物样品中分离的培养的菌株的反射(阿诺德等人,2001;希金斯等人,2007; Terhonen 等人,2011)。使用商业试剂盒处理大量样品进行常规PCR反应的成本可能很高,此外,对2-巯基乙醇,氯仿或苯酚等有害化学品的需求也可能增加。限制协议在教学和培训中的适用性。

...

Assessment of Caenorhabditis elegans Competitive Fitness in the Presence of a Bacterial Parasite
Author:
Date:
2018-08-20
[Abstract]  Accurate measurements of an organism’s fitness are crucial for measuring evolutionary change. Methods of fitness measurement are most accurate when incorporating an individual’s survival and fecundity, as well as accounting for any ecological interactions or environmental effects experienced by the organism. Here, we describe a protocol for measuring the relative mean fitness of Caenorhabditis elegans populations, or strains, through an assay that accounts for individual survival, fecundity, and intraspecific competitive ability in the presence of a bacterial parasite. In this competitive fitness assay nematodes from a focal population or strain are mixed with a GFP-marked tester strain in equal proportions, the mixture of nematodes are then exposed to a parasite, and the ... [摘要]  准确测量生物体的适应度对于衡量进化变化至关重要。当结合个体的生存和繁殖力以及考虑生物体所经历的任何生态相互作用或环境影响时,适合度测量的方法是最准确的。在这里,我们描述了一种用于测量秀丽隐杆线虫种群或菌株的相对平均适合度的方案,该方案通过在细菌寄生虫存在下计算个体存活率,繁殖力和种内竞争能力的测定。在这种竞争性适应性测定中,来自焦点种群或菌株的线虫与GFP标记的测试菌株以相等比例混合,然后将线虫的混合物暴露于寄生虫,并且通过测量来确定焦点菌株的相对竞争适合度。一代后,焦点线虫与GFP标记的线虫的比例发生变化。具体地,可以实施该协议以通过确定进化线虫与祖先线虫群体的相对竞争适合度来测量实验进化后线虫宿主适应度的变化。

【背景】 准确测量适应度和适应度随时间的变化对于确定群体对自然选择的反应至关重要。尽管如此,健康是众所周知难以衡量的,因为它包含了个人的生存,繁殖力,繁殖时间,并且必须考虑到对个体的生态和环境影响。尽管在所有可能的条件下没有用于测量适合度的协议是最佳的,但是考虑到生存和繁殖力的适应度的测量,同时保持生态和环境影响不变,可能提供对给定场景的适合性的可靠总体估计。在这里,我们描述了一个用于测量 C之间相对适应度差异的协议。线虫种群或菌株,用于确定细菌寄生虫存在时进化时间内相对适合度的变化。我们利用革兰氏阴性细菌粘质沙雷氏菌作为 ...

Two Different Methods of Quantification of Oxidized Nicotinamide Adenine Dinucleotide (NAD+) and Reduced Nicotinamide Adenine Dinucleotide (NADH) Intracellular Levels: Enzymatic Coupled Cycling Assay and Ultra-performance Liquid Chromatography (UPLC)-Mass Spectrometry
Author:
Date:
2018-07-20
[Abstract]  Current studies on the age-related development of metabolic dysfunction and frailty are each day in more evidence. It is known, as aging progresses, nicotinamide adenine dinucleotide (NAD+) levels decrease in an expected physiological process. Recent studies have shown that a reduction in NAD+ is a key factor for the development of age-associated metabolic decline. Increased NAD+ levels in vivo results in activation of pro-longevity and health span-related factors. Also, it improves several physiological and metabolic parameters of aging, including muscle function, exercise capacity, glucose tolerance, and cardiac function in mouse models of natural and accelerated aging.

Given the importance of monitoring cellular NAD+ and ...
[摘要]  目前关于代谢功能障碍和虚弱的年龄相关发展的研究每天都有更多的证据。众所周知,随着衰老的进展,烟酰胺腺嘌呤二核苷酸(NAD + )水平在预期的生理过程中降低。最近的研究表明,NAD + 的减少是与年龄相关的代谢衰退发展的关键因素。增加NAD + 水平体内导致激活寿命和健康跨度相关因素。此外,它改善了老化的几个生理和代谢参数,包括自然和加速老化的小鼠模型中的肌肉功能,运动能力,葡萄糖耐量和心脏功能。

鉴于监测细胞NAD + 和NADH水平的重要性,有一个值得信赖的方法是至关重要的。该方案的目的是在有效且广泛适用的测定中描述来自组织和细胞的NAD + 和NADH提取以及其图形和定量分析。

【背景】氧化烟酰胺腺嘌呤二核苷酸和还原型烟酰胺腺嘌呤二核苷酸(NAD + 和NADH)是重要的生物辅助因子,它们在几种合成代谢和分解代谢功能中提供和接受电子。它们参与诸如糖酵解,三羧酸循环和氧化磷酸化的反应。此外,它还作为DNA损伤修复中涉及的几种酶的底物,例如sirtuins和poly(ADP-核糖)聚合酶(PARP)(Imai和Guarente,2014; Verdin,2015; Yoshino et al。 ,2018)。

NAD + ...

Comments