NR-19744炭疽芽孢杆菌Gateway?克隆套装,在大肠杆菌中重组,板20(克隆)
The Bacillus anthracis (B. anthracis) Gateway? clone set consists of 58 plates which contain 5341 sequence validated clones from B. anthracis, strains Ames (5139 clones), Sterne (107 clones; contains plasmid pXO1 only) and A2012 (95 clones; contains plasmid pXO2 only) cloned in Escherichia coli (E. coli) DH10B-T1 cells. Each open reading frame was constructed in vector pDONR?221 (Invitrogen?) with a native start codon and no stop codon. The library was independently cloned and sequence verified by the Harvard Institute of Proteomics.
Information related to the use of Gateway? Clones can be obtained from Invitrogen?. Recombination was facilitated through a Harvard-modified attB substrate (attB-PCR product or a linearized attB expression clone) with an attP substrate (pDONR?221) to create an attL-containing entry clone. The entry clone contains recombinational cloning sites, attL1 and attL2 to facilitate gene transfer into a destination vector, M13 forward and reverse priming sites for sequencing and a kanamycin resistance gene for selection. Please refer to the Invitrogen? Gateway? Technology Manual for additional details.
Plate orientation and viability were confirmed for NR-19744.
Each inoculated well of the 96-well plate contains approximately 60 ?L of E. coli culture (strain DH10B-T1) in Luria Bertani (LB) broth containing 50 ?g/mL kanamycin supplemented with 15% glycerol.
Clone plates are replicated using a BioMek? FX robot. Production in the 96-well format has increased risk of cross-contamination between adjacent wells. Individual clones should be purified (e.g. single colony isolation and purification using good microbiological practices) and sequence-verified prior to use. BEI Resources only confirms the clone plate orientation and viability of randomly picked clones. BEI Resources does not confirm or validate individual clone identities provided by the contributor.
BEI Resources 由美国国家过敏和传染病研究所 ( NIAID )成立,旨在为研究 A、B 和 C 类优先病原体、新兴传染病病原体、非病原微生物和其他相关微生物材料提供试剂、工具和信息到研究界。BEI Resources 获取、验证和生产科学家进行基础研究和开发改进的诊断测试、疫苗和疗法所需的试剂。通过将这些功能集中在 BEI Resources 中,可以监控科学界对这些材料的访问和使用,并确保试剂的质量控制。
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BEI Resources 自 2003 年起由美国典型培养物保藏中心 (ATCC) 根据合同管理。2016 年 5 月, ATCC获得了一份为期七年的继续管理 BEI Resources的合同。合同范围已扩大到更全面的研究目录材料,包括由其他政府支持的研究项目存放的材料,将提供给生物防御和新兴传染病科学界。真菌、寄生虫、载体和其他相关材料已添加到现有的细菌、病毒和毒素试剂中,涵盖 NIAID A、B 和 C 类优先病原体和 NIAID 指定的新发传染病病原体和生物。