One of the major problems in the U.S. and global cotton production is the damage caused by the reniform nematode, Rotylenchulus reniformis. Amplification of DNA from single nematodes for further molecular analysis can be challenging sometimes. In this research, two whole-genome amplification (WGA) methods were evaluated for their efficiencies in DNA amplification from a single reniform nematode. The WGA was carried out using both REPLI-g Mini and Midi kits, and the GenomePlex single cell whole-genome amplification kit. Sequence analysis produced 4 Mb and 12 Mb of genomic sequences for the reniform nematode using REPLI-g and SIGMA libraries. These sequences were assembled into 28,784 and 24,508 contigs, respectively, for REPLI-g and SIGMA libraries. The highest repeats in both libraries were of low complexity, and the lowest for the REPLI-g library were for satellites and for the SIGMA library, RTE/BOV-B. The same kind of repeats were observed for both libraries; however, the SIGMA library had four other repeat elements (Penelope (long interspersed nucleotide element (LINE)), RTE/BOV-B (LINE), PiggyBac, and Mirage/P-element/Transib), which were not seen in the REPLI-g library. DNA transposons were also found in both libraries. Both reniform nematode 18S rRNA variants (RN_VAR1 and RN_VAR2) could easily be identified in both libraries. This research has therefore demonstrated the ability of using both WGA methods, in amplification of gDNA isolated from single reniform nematodes.

中文翻译：

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表征肾形线虫基因组中两种全基因组扩增方法中的重复序列

美国乃至全球棉花生产的主要问题之一是肾形线虫轮状线虫（Roylylulus reniformis）造成的损害。。从单个线虫中扩增DNA进行进一步的分子分析有时可能具有挑战性。在这项研究中，评估了两种全基因组扩增（WGA）方法从单个肾形线虫进行DNA扩增的效率。使用REPLI-g Mini和Midi试剂盒以及GenomePlex单细胞全基因组扩增试剂盒进行WGA。序列分析使用REPLI-g和SIGMA文库为肾形线虫产生了4 Mb和12 Mb的基因组序列。对于REPLI-g和SIGMA文库，这些序列分别被组装成28,784和24,508个重叠群。这两个库中的最高重复序列的复杂性较低，而REPLI-g库的最低重复序列则是卫星，而SIGMA库的RTE / BOV-B则最低。在两个文库中都观察到相同类型的重复；然而，SIGMA文库中还有其他四个重复元件（佩内洛普（长穿插核苷酸元件（LINE）），RTE / BOV-B（LINE），PiggyBac和Mirage / P元素/ Transib），这在REPLI-g中是看不到的图书馆。在两个文库中也发现了DNA转座子。可以很容易地在两个文库中识别出两个肾形线虫18S rRNA变体（RN_VAR1和RN_VAR2）。因此，这项研究证明了使用两种WGA方法扩增从单个肾形线虫分离的gDNA的能力。可以很容易地在两个文库中识别出两个肾形线虫18S rRNA变体（RN_VAR1和RN_VAR2）。因此，这项研究证明了使用两种WGA方法扩增从单个肾形线虫分离的gDNA的能力。可以很容易地在两个文库中识别出两个肾形线虫18S rRNA变体（RN_VAR1和RN_VAR2）。因此，这项研究证明了使用两种WGA方法扩增从单个肾形线虫分离的gDNA的能力。