博凯森生物利用PacBio Sequel系统提供真菌全基因组测序，以提供更多关于遗传结构和功能的见解。我们在知识、实践和经验方面都是最好的。
真菌全基因组介绍 重新 测序服务
Fungi play vital roles in the biosphere and are closely related to human life with important medical and economic value. Fungi are involved in a wide range of activities—some fungi are decomposers, parasites or pathogens of other organisms, and others are beneficial partners in symbiosis with animals, plants or algae. Some fungi affect human health in various ways. Fungi are also the major sources of antibiotics, such as penicillin from the fungus 青霉菌。
Obtaining fungal whole genome sequence is the basics of fungi research, and to better understand fungal biodiversity, growth, nutrition, physiology, genetics, metabolism, and ecology. Thus, hundreds of fungal genomes have been sequenced and are publically available today with a significant increase, although these initiatives have typically yielded considerably fragmented genome assemblies, they often lack large contiguous genomic regions. Many important genomic features are contained in intergenic DNA that is often missing in current genome assemblies.
To solve this problem, 博凯森生物 introduces the PacBio platform. This machine uses single molecule real-time (SMRT) detection technology that achieves real-time sequencing of individual polymerase molecules. SMRT detection is based on the properties of zero-mode waveguides (ZMWs), consisting of DNA polymerases bound to nanophotonic confinement structures. This technology does not require amplification of the genomic DNA, which addresses one of the major problems of second generation sequencing technologies; thus leading to the least degree of bias and longer read lengths (average >15,000 bp, some reads >100,000 bp). With the long reads of SMRT Sequencing on the PacBio System, we can generate complete 重新 assemblies for fungal genomes achieve or approximate 0 gaps or N-based errors, contig N50＞1 Mb, 99.999% accuracy. Additionally, we provide 微生物全基因组测序 通过使用下一代测序。
项目工作流程
我们经验丰富的专主页团队通过遵循每一个程序来执行质量管理，以确保全面准确的结果。真菌全基因组测序的一般工作流程概述如下。
样品要求：
DNA amount: ≥ 10 μg
DNA纯度：OD260/280=1.8~2.0，无降解或RNA污染
测序服务策略：
20 kb Library, ≥ 60X genome coverage depth
数据分析
- 基因组组装
- 基因预测
- 基因注释
- 比较物种基因组分析
分析管道
PacBio是第三代测序仪，具有高度的鲁棒性和成本效益，应该是真菌基因组测序的首选平台，特别是对于那些众所周知难以测序的真菌。 博凯森生物 has extensive experience to offer the fungi whole genome sequencing service. Please contact us for more information and a detailed quote.
1.真菌样品制备面临哪些挑战？
一些样本容易降解，因此我们建议在DNA提取后快速构建文库并测序。一些真菌菌株很难培养，因此建议使用少量DNA或全基因组扩增构建文库。对于难以分离DNA的样本，您可以采用文献中的方法。
2.为什么需要真菌基因组调查？
与细菌相比，真菌基因组、质粒和样品制备更为复杂。此外，只有少数真菌基因组被发表。通过基因组调查，我们可以获得基因组大小、GC含量、重复序列和质粒的信息，这可以为实质性测序和基因组组装铺平道路。
两个基因组的序列、组装和鉴定 果甲草 用作采后病害生物防治剂的菌株
背景
酵母 果甲草 was reported as an efficient biological control agent of postharvest diseases of fruits and vegetables, and it is the bases of the commercial formulated product “Shemer.” Researchers assembled the whole genome sequence of two strains of 果锥分枝杆菌 .
结果菌株277 果锥分枝杆菌 在PacBio RS II测序仪（P6-C4试剂盒，20 Kb文库，24个SMRT细胞，靶覆盖率为20X）上测序，得到由93个重叠群组成的高质量基因组草案，N50为957836 bp。估计的基因组大小约为26 Mb。用MAKER预测了8629个基因，用Blast2GO和InterProScan成功注释了6262个基因。
果锥分枝杆菌 AP47采用Illumina MiSeq技术进行测序，文库插入长度为330 bp，配对末端为300 bp。两者的基因组大小（26Mb） 果锥分枝杆菌 菌株以及突变率可能表明 果锥分枝杆菌 可以经历基因组变化以适应植物表面、耐受各种环境胁迫并在有限的营养资源下生存。
参考：
Edoardo P。； 等两种基因的序列、组装和鉴定 果甲草 用作采后病害生物防治剂的菌株。 微生物学前沿. 2017, 8:1-15.
