自十年前第二代测序技术(NGS)进入市场以来,发展迅猛。测序速度提高了,通量明显攀升,费用却在降低,成为了许多生命科学研究者的常规手段。许多科学家把他们第一次使用NGS产生的疑问反映给了MOBIO技术部。其中最常见的问题是使用某款MO BIO 的提取试剂盒所提到的DNA能否直接用于第二代测序。
他们在担心什么呢?因为成功的NGS要求从样品中提取的DNA不能含有盐、蛋白、EDTA等其它难缠的杂质。如果要处理的是"复杂"样品,像我们大多数客户所做的,这确实是个问题。这类复杂样品包括:鸟粪、昆虫肠内容物、仙人掌肉、管道粘液、泥炭苔、养猪场污水、废水,乃至原油污染的土壤(听起来就像电视剧《Dirty Jobs》中的桥段,要从这些样品中获得干净的DNA还是极具挑战的。但也不是没有可能。我们的许多客户已经在使用MOBIO的试剂盒提取DNA直接用于第二代测序,有些则用MOBIO的PowerClean DNA Clean-up Kit去除DNA粗品的杂质使其达到测序使用要求。IRT,我们的专利抑制因子去除技术(Inhibitor Removal Technology),能非常高效去除影响第二代测序的所有杂质。
有客户向我们索取参考文献。我们在下面列了个简表,尽可能涵盖各种样品类型、不同第二代测序平台和所使用的MO BIO提取试剂盒。希望这些能有助于你的研究以及第二代测序。如果你有其他相关的参考文献也可以告诉我们。
Virus, UltraClean Microcial DNA Isolation Kit, Roche 454 Pyrosequencing
Metagenomic Analysis of the Viral Communities in Fermented Foods
(发酵食物中病毒群落元基因组学分析,罗氏454焦磷酸测序)
Eun-Jin Park, Kyoung-Ho Kim, Guy C. J. Abell, Min-Soo Kim, Seong Woon Roh, and Jin-Woo Bae. Appl. Envir. Microbiol., Feb 2011; 77: 1284 – 1291.
Bacteria, UltraClean Microbial DNA Isolation Kit, Pyrosequencing
Genome Sequence of a Novel Species, Propionibacterium humerusii
(新细菌物种基因组序列,焦磷酸测序)
Susan M. Butler-Wu, Dhruba J. Sengupta, Weerayuth Kittichotirat, Frederick A. Matsen, III, and Roger E. Bumgarner J. Bacteriol., Jul 2011; 193: 3678
DNA, PowerClean DNA Clean-Up Kit, Pyrosequencing
Lachnospiraceae and Bacteroidales Alternative Fecal Indicators Reveal Chronic Human Sewage Contamination in an Urban Harbor
(选择毛螺菌科或拟杆菌目作为长期受生活污水污染城市港口粪便污染指示菌)
Ryan J. Newton, Jessica L. VandeWalle, Mark A. Borchardt, Marc H. Gorelick, and Sandra L. McLellan. Appl. Envir. Microbiol., Oct 2011; 77: 6972 – 6981.
Deep Sea Sediments, PowerMax DNA Isolation Kit, Genome Analyzer GAII instrument (Illumina)
Ultra-deep sequencing of foraminiferal microbarcodes unveils hidden richness of early monothalamous lineages in deep-sea sediments.
Beatrice Lecroq, Franck Lejzerowicz, Dipankar Bachar, Richard Christen, Philippe Esling, Loic Baerlocher, Magne Osteras, Laurent Farinelli, and Jan Pawlowski. PNAS, Aug 2011; 108: 13177 – 13182.
Symbiotic-fungus, PowerSoil DNA Isolation Kit, Pyrosequencing
Distinct Ectomycorrhizospheres Share Similar Bacterial Communities as Revealed by Pyrosequencing-Based Analysis of 16S rRNA Genes
(焦磷酸测序分析16S rRNA 基因表明不同Ectomycorrhizospheres有类似的细菌群落多样性)
S. Uroz, P. Oger, E. Morin, and P. Frey-Klett. Appl. Envir. Microbiol., Apr 2012; 78: 3020 – 3024.
Metal-Contaminated Stream Sediment, PowerSoil DNA Isolation Kit, GS 454 FLX Pyrosequencing
Mercury and Other Heavy Metals Influence Bacterial Community Structure in Contaminated Tennessee Streams
(水银及其它重金属污染对田纳西溪流沉积物中细菌群落结构的影响)
Tatiana A. Vishnivetskaya, Jennifer J. Mosher, Anthony V. Palumbo, Zamin K. Yang, Mircea Podar, Steven D. Brown, Scott C. Brooks, Baohua Gu, George R. Southworth, Meghan M. Drake, Craig C. Brandt, and Dwayne A. Elias, Appl. Envir. Microbiol., Jan 2011; 77: 302 – 311.
Skin Swabs, PowerSoil DNA Isolation Kit, Pyrosequencing
Forensic identification using skin bacterial communities
(皮肤微生物群落分析作法医鉴定)
Noah Fierer, Christian L. Lauber, Nick Zhou, Daniel McDonald, Elizabeth K. Costello, and Rob Knight PNAS, Apr 2010; 107: 6477 – 6481.
Oral swabs, PowerSoil DNA Isolation Kit, 454 Life Sciences FLX Sequencer
The bacterial microbiota in the oral mucosa of rural Amerindians
(美洲印第安人口腔棉拭子微生物)
Monica Contreras, Elizabeth K. Costello, Glida Hidalgo, Magda Magris, Rob Knight, and Maria G. Dominguez-Bello, Microbiology, Nov 2010; 156: 3282 – 3287.
Breast Tissue, FFPE DNA Isolation Kit, SOLiD Sequencing
Identification of high-confidence somatic mutations in whole genome sequence of formalin-fixed breast cancer specimens
(福尔马林固定乳房癌组织全基因组序列中识别高可信度体细胞突变)
Yost SE, Smith EN, Schwab RB, Bao L, Jung H, Wang X, Voest E, Pierce JP, Messer K, Parker BA, Harismendy O, Frazer KA Nucleic Acids Res. 2012 Apr 6, 1-12
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提到参考文献,MOBIO有一个参考文献计划,如果你在你们文章中提到MOBIO试剂盒,你将有机会得到我们送的免费试剂盒。
感谢您的阅读!