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产品图片
货号/SKU
NR-19785
货号/规格
EA
库存与交货期
3-8周
人民币价格
14000
试剂海关审批
A/B级风险物质只能直接使用人购买并持有实验室有效资质，其它询客服确认
国外采购
支持/部分限制一年内购买数量
厂牌
BEI Resources(ATCC)
产品基础信息
生物安全等级建议分类：美国、1
产品描述信息
NR-19785?? Mycobacterium tuberculosis, Strain CDC1551, Knockout Gateway? Clone Set, Recombinant in Escherichia coli, Plate 3(Clones)|Mycobacterium tuberculosis|Mycobacterium tuberculosis, Strain CDC1551, Knockout Gateway? Clone Set, Recombinant in Escherichia coli, Plate 3|-80°C or colder|Pathogen Functional Genomics Resource Center at the J. Craig Venter InstituteAcknowledgment for publications should read "The following reagent was obtained through BEI Resources, NIAID, NIH: Mycobacterium tuberculosis, Strain CDC1551, Knockout Gateway? Clone Set, Recombinant in Escherichia coli, Plate 3, NR-19785."|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), Knockout Gateway? clone set consists of 8 plates which contain 641 sequence validated knockout clones from M. tuberculosis, strain CDC1551. Each open reading frame was constructed with a hygromycin selectable gene replacement marker in vector pDEST-YUB, a Gateway? compatible adaptation of the cosmid cloning vector pYUB854 and cloned in Escherichia coli (E. coli) DH10B-T1 cells. The final construct also contains the ?-lactamase gene to confer ampicillin resistance for plasmid selection in E. coli. The sequence was validated by full length sequencing of each 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?. A PCR product representing a functional hygromycin resistance cassette was assembled with chromosomal amplicons of approximately 600 base pairs of the regions flanking each gene targeted for replacement. The three fragments (left flank, hygromycin resistance gene, right flank) were amplified and cloned into pDONR? entry vectors (Invitrogen?). Recombination was facilitated through an attB substrate (attB-PCR product or a linearized attB expression clone) with an attP substrate (pDONR? vector) to create an attL-containing entry clone using the three-fragment MultiSite Gateway? Pro method. The hygromycin resistance cassette was sequence verified and experimentally verified through hygromycin resistance of DH10B-T1 E. coli cells. The final destination construct was confirmed by restriction digestion analysis. Please refer to the Invitrogen? Gateway? Technology Manual for additional Gateway? product details.
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 100 ?g/mL ampicillin supplemented with 15% glycerol.
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.
The Mycobacterium tuberculosis (M. tuberculosis), Knockout Gateway? clone set consists of 8 plates which contain 641 sequence validated knockout clones from M. tuberculosis, strain CDC1551. Each open reading frame was constructed with a hygromycin selectable gene replacement marker in vector pDEST-YUB, a Gateway? compatible adaptation of the cosmid cloning vector pYUB854 and cloned in Escherichia coli (E. coli) DH10B-T1 cells. The final construct also contains the ?-lactamase gene to confer ampicillin resistance for plasmid selection in E. coli. The sequence was validated by full length sequencing of each 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?. A PCR product representing a functional hygromycin resistance cassette was assembled with chromosomal amplicons of approximately 600 base pairs of the regions flanking each gene targeted for replacement. The three fragments (left flank, hygromycin resistance gene, right flank) were amplified and cloned into pDONR? entry vectors (Invitrogen?). Recombination was facilitated through an attB substrate (attB-PCR product or a linearized attB expression clone) with an attP substrate (pDONR? vector) to create an attL-containing entry clone using the three-fragment MultiSite Gateway? Pro method. The hygromycin resistance cassette was sequence verified and experimentally verified through hygromycin resistance of DH10B-T1 E. coli cells. The final destination construct was confirmed by restriction digestion analysis. Please refer to the Invitrogen? Gateway? Technology Manual for additional Gateway? product details.
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 100 ?g/mL ampicillin supplemented with 15% glycerol.
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.
主要内容
此项目的每个订单数量限制为1.此商品每年可订购两次.通过此限制的订单将在发货前发送到NIAID进行批准. 结核分枝杆菌（M.Tuberculosis），敲除网关?克隆集由8个板组成，其中包含641个序列验证的敲除克隆来自 m.结核病，菌株CDC1551.每个开放阅读框架用卷曲霉素选择基因替代标记在载体pdest-yub中构建，网关?兼容的粘性克隆载体pyub854的适应性，并克隆在大肠杆菌（大肠杆菌）中 DH10B-T1细胞.最终的构建体还含有β-内酰胺酶基因，以赋予氨苄青霉素的抗性抗质粒选择. Coli .通过大于1x覆盖率的每个克隆的全长测序验证序列，突变率小于0.2％. 与使用网关?克隆有关的信息从 Invitrogen ?.代表功能性潮霉素抵抗盒的PCR产物用染色区域的染色体扩增器组装，每个基因靶向更换.将三个片段（左侧侧翼，潮霉素抵抗基因，右侧）扩增并克隆到PDONR TM进入载体（Invitrogen TM）中.通过 att b底板（ att b-pCR产物或线性化的 att b表达克隆）促进重组，用 att P衬底（PDONR?矢量）使用三个片段多路网关? pro 方法.潮霉素抵抗盒是序列验证和通过DH10B-T1的潮霉素抗性进行了实验验证. Coli 细胞.通过限制消化分析证实了最终目的地构建体.请参阅Invitrogen?网关?技术手册用于额外的网关?产品详细信息. 96孔板的每个接种井含有大约60μl的 e. Coli 培养（菌株DH10B-T1）在含有100μg/ ml氨苄青霉素的LURIA Bertani（LB）肉汤中，补充有15％甘油. 生产在96中 - 相邻井之间的交叉污染风险增加了.应纯化各个克隆（例如，使用良好的微生物实践的单菌落分离和纯化）并在使用前进行序列验证. Bei资源不确认或验证贡献者提供的个体突变体.
厂牌介绍
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。
除了为传染病界提供材料外，BEI Resources 还鼓励和支持研究人员和机构的材料存放。使用 BEI Resources存放材料对研究人员和研究社区有许多优势，包括安全存储、社区访问和分发；同时保护存款人的知识产权。只要有需要，BEI 资源库将作为研究人员的资源进行维护。您在 BEI Resources 的存款是一项有助于未来研究的长期投资。
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。
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