NR-19661_结核分枝杆菌Gateway?克隆套装,在大肠杆菌中重组,板25(克隆)
Quantity limit per order for this item is 1. This item can be ordered twice a year. Orders over this limit will be sent to NIAID for approval before shipment.
The Mycobacterium tuberculosis (M. tuberculosis) Gateway? clone set consists of 42 plates which contain 3724 sequence validated clones (3294 M. tuberculosis, strain H37Rv clones supplemented with 430 unique open reading frames (ORF) from M. tuberculosis, strain CDC1551) cloned in Escherichia coli (E. coli) DH10B-T1 cells. Each ORF was recombined in vector pDONR?221 with an ATG start codon and no stop codon. The sequence was validated by full length sequencing of each entry clone with greater than 1X coverage and a mutation rate of less than 0.2%.
Information related to the use of Gateway? Clones can be obtained from Invitrogen?. Recombination was facilitated through an 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.
Each 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.
Note: 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 does not confirm or validate individual mutants provided by the contributor.
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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。