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illustra Nucleon PhytoPure

插图Nucleon PhytoPure

Company: Cytiva
Catalog#: RPN8510
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Chromosome Dosage Analysis in Plants Using Whole Genome Sequencing
[Abstract]  Relative chromosome dosage, i.e., increases or decreases in the number of copies of specific chromosome regions in one sample versus another, can be determined using aligned read-counts from Illumina sequencing (Henry et al., 2010). The following protocol was used to identify the different classes of aneuploids that result from uniparental genome elimination in Arabidopsis thaliana, including chromosomes that have undergone chromothripsis (Tan et al., 2015). Uniparental genome elimination results in the production of haploid progeny from crosses to specific strains called “haploid inducers” (Ravi et al., 2014). On the other hand, chromothripsis, which was first discovered in cancer genomes, is a phenomenon that results in clustered, ... [摘要]  可以使用来自Illumina测序的对准读数确定相对染色体剂量,即,一个样品中特定染色体区域的拷贝数相对于另一个样品的拷贝数的增加或减少(Henry等人,2010)。以下方案用于鉴定拟南芥中单亲基因组消除的不同类型的非整倍体,包括染色体发生染色体(Tan等,2015)。单亲基因组消除导致从杂交到称为“单倍体诱导物”的特异性菌株的单倍体后代的产生(Ravi等,2014)。另一方面,在癌症基因组中首次发现的chromothripsis是导致聚类,高度重排的染色体的现象。在植物中,作为基因组消除的结果已经观察到chromothripsis(Tan等人,2015)。检测染色体剂量的变化在与基因组消除相关的那些旁边有多种应用。例如,通过花粉粒的γ-照射产生杨树杂种的剂量变异种群。使用该技术鉴定了数百个剂量损伤,插入和缺失,并提供了一种将基因座与在该群体中观察到的表型后果相关联的方法(Henry等,2015)。
该方法已成功应用于许多不同物种的染色体用量变化,包括拟南芥(Tan et al。,2015),拟南芥(Ravi et al。,2014),水稻(Henry et ...

DNA Fragmentation Analysis
[Abstract]  DNA fragmentation with length corresponding to multiple integer of approximately 180 base pairs is a distinct feature of apoptosis in animals and programmed cell death in plants. This feature can simply be detected by DNA gel electrophoresis followed by ethidium bromide staining, although in some cases it is difficult to distinguish the DNA laddering. We herein describe a protocol to detect a programmed cell death-associated DNA laddering of plant tissues. After agarose-gel electrophoresis of genomic DNA, Southern hybridization using DIG-labeled genomic DNA probe is performed, that improves detection of DNA laddering. [摘要]  具有对应于大约180个碱基对的多个整数的长度的DNA片段是动物中凋亡和植物中程序性细胞死亡的独特特征。 这个特征可以简单地通过DNA凝胶电泳随后溴化乙锭染色检测,尽管在一些情况下难以区分DNA梯状。 我们在本文中描述了用于检测植物组织的程序性细胞死亡相关DNA梯状的方案。 在基因组DNA的琼脂糖凝胶电泳后,进行使用DIG标记的基因组DNA探针的Southern杂交,其改善DNA梯状的检测。