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Spermidine

亚精胺

Company: Sigma-Aldrich
Catalog#: S0266
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Pollen Germination and Pollen Tube Growth of Arabidopsis thaliana: In vitro and Semi in vivo Methods
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Date:
2018-08-20
[Abstract]  Studies of pollen germination and post-germination development are not only essential for understanding plant reproduction but also are an excellent model system for tip-based growth. Here we describe easy, reproducible methods for germination and growth of pollen from the model plant Arabidopsis thaliana in artificial conditions. Our growth system can be used both for pollen placed directly on this artificial substrate as well as for the so-called ‘semi in vivo’ method. This is where a pistil is cut shortly after hand-pollination and the pollen tubes grow through the plant tissue and emerge from the cut end onto the surface of the artificial medium. [摘要]  花粉萌发和萌发后发育的研究不仅对于理解植物繁殖是必不可少的,而且也是用于尖端生长的优秀模型系统。 在这里,我们描述了简单,可重复的方法,用于在人工条件下从模式植物拟南芥中萌发和生长花粉。 我们的生长系统既可用于直接置于该人工基质上的花粉,也可用于所谓的“半体内”方法。 这是在手动授粉后不久切割雌蕊并且花粉管通过植物组织生长并从切割末端出现在人工培养基表面上的地方。

【背景】 开花植物的花粉被广泛用作快速,尖端生长的模型系统。当然,花粉生物学的研究对于了解植物的生育力和生殖发育也是必不可少的。然而,一种简单可靠的方法是从模式植物拟南芥中萌发花粉并在体外维持快速,形态正常的花粉管生长令人沮丧地难以捉摸。在开发我们自己的方法(Rodriguez-Enriquez et al。,2013)之前,我们尝试复制几种已发表的方法,但经过多次尝试后,我们未能产生令人满意的结果。例如,在实验室中复制Boavida和McCormick的方法很困难,因为它具有非常窄的温度最佳值(22°C),这种情况显然不能反映 A的生殖生物学。拟南芥(Boavida和McCormick,2007)。此外,在该方法中存在发芽率和局部花粉密度的问题,其也不完全反映在柱头表面上的自然事件。我们的第一个线索是,体外可靠的花粉萌发所需的关键“缺失因子”来自观察到放置 ...

Genome Editing in Diatoms Using CRISPR-Cas to Induce Precise Bi-allelic Deletions
Author:
Date:
2017-12-05
[Abstract]  Genome editing in diatoms has recently been established for the model species Phaeodactylum tricornutum and Thalassiosira pseudonana. The present protocol, although developed for T. pseudonana, can be modified to edit any diatom genome as we utilize the flexible, modular Golden Gate cloning system. The main steps include how to design a construct using Golden Gate cloning for targeting two sites, allowing a precise deletion to be introduced into the target gene. The transformation protocol is explained, as are the methods for screening using band shift assay and/or restriction site loss. [摘要]  最近为三角褐指藻(Phaeodactylum tricornutum)和海绵假丝酵母(Thalassiosira pseudonana)建立了硅藻基因组编辑。 目前的协议,虽然开发的 T。 pseudonana ,可以修改编辑任何硅藻基因组,因为我们利用灵活,模块化的金门克隆系统。 主要步骤包括如何设计构建使用金门克隆靶向两个网站,允许一个精确的删除被引入目标基因。 解释转化方案,以及使用带移位测定和/或限制性位点丢失进行筛选的方法。

【背景】CRISPR-Cas正在迅速成为分子研究的一个关键方法。基于在细菌和古细菌中发现的病毒防御机制,CRISPR-Cas诱导基因组中精确位置的双链断裂(DSBs)。它涉及使用与CRISPR ...

MNase Digestion for Nucleosome Mapping in Neurospora
Author:
Date:
2016-06-05
[Abstract]  Digestion of chromatin by micrococcal nuclease MNase followed by high throughput sequencing allows us to determine the location and occupancy of nucleosomes on the genome. Here in this protocol we have described optimized conditions of MNase digestion of filamentous fungus Neurospora crassa chromatin without a requirement of a nuclear fractionation step. [摘要]  通过微球菌核酸酶MNase消化染色质,然后高通量测序允许我们确定核小体在基因组上的位置和占据。 在这个协议中,我们描述了MNase消化丝状真菌粗糙链孢霉染色体的优化条件,而不需要核分馏步骤。

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