NR-19437_烟曲霉,KnockoutGateway?克隆套装,在大肠杆菌中重组(克隆）
产品图片
货号/SKU
NR-19437
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-19437?? Aspergillus fumigatus, Knockout Gateway? Clone Set, Recombinant in Escherichia coli(Clones)|Aspergillus fumigatus|Aspergillus fumigatus, Knockout Gateway? Clone Set, Recombinant in Escherichia coli|-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: Aspergillus fumigatus, Knockout Gateway? Clone Set, Recombinant in Escherichia coli, NR-19437."|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 Aspergillus fumigatus (A. fumigatus), Gateway? clone set consists of 1 plate which contains 70 sequence validated knockout clones from A. fumigatus, strain A1163 with ampicillin selectable pyrG selection gene, cloned in Escherichia coli (E. coli) DH10B-T1 cells. Each open reading frame was constructed in vector pDEST? 17 (Invitrogen?). Successful cloning was determined using a restriction digestion approach.
Information related to the use of Gateway? Clones can be obtained from Invitrogen?. Using the three fragment MultiSite Gateway? strategy the pyrG selectable marker was assembled with chromosomal amplicons (~ 1Kb) of the regions flanking the targeted gene. The three fragments (left flank, pyrG, right flank) were amplified and cloned into entry vector pDONR?221 P1-P4, pDONR?221 P4r-P3r and pDONR?221 P3-P2, respectively. The flanking clones were confirmed by PCR, the pyrG clone was sequence verified. The final assembled construct is created using LR Clonase? II Plus to recombine the three clones with each other and the destination vector, pET-DEST-TIGR02. Linearized vector can be used to transform chemically competent protoplasts. Upon homologous recombination the pyrG gene complements an auxotrophic mutant A1163 strain. Please refer to the Invitrogen? Gateway? Technology Manual for additional details.
Plate orientation and viability were confirmed for NR-19437.
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 ampicilln 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.
The Aspergillus fumigatus (A. fumigatus), Gateway? clone set consists of 1 plate which contains 70 sequence validated knockout clones from A. fumigatus, strain A1163 with ampicillin selectable pyrG selection gene, cloned in Escherichia coli (E. coli) DH10B-T1 cells. Each open reading frame was constructed in vector pDEST? 17 (Invitrogen?). Successful cloning was determined using a restriction digestion approach.
Information related to the use of Gateway? Clones can be obtained from Invitrogen?. Using the three fragment MultiSite Gateway? strategy the pyrG selectable marker was assembled with chromosomal amplicons (~ 1Kb) of the regions flanking the targeted gene. The three fragments (left flank, pyrG, right flank) were amplified and cloned into entry vector pDONR?221 P1-P4, pDONR?221 P4r-P3r and pDONR?221 P3-P2, respectively. The flanking clones were confirmed by PCR, the pyrG clone was sequence verified. The final assembled construct is created using LR Clonase? II Plus to recombine the three clones with each other and the destination vector, pET-DEST-TIGR02. Linearized vector can be used to transform chemically competent protoplasts. Upon homologous recombination the pyrG gene complements an auxotrophic mutant A1163 strain. Please refer to the Invitrogen? Gateway? Technology Manual for additional details.
Plate orientation and viability were confirmed for NR-19437.
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 ampicilln 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.
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. 曲霉菌fumigatus（A. fumigatus），网关?克隆设置由1个板组成，其中包含来自 a的70个序列验证的敲除克隆. Fumigatus ，菌株A1163具有氨苄青霉素选择的Pyrg选择基因，克隆在大肠杆菌（大肠杆菌）/ I> DH10B-T1细胞中.每个开放阅读框架是在传染媒介 pdest?17 （Invitrogen?）中.使用限制消化方法确定成功的克隆. 与网关使用的信息相关?克隆可以从?.使用三个片段多态网关?策略，Pyrg可选标记与侧翼靶向基因的区域的染色体扩增子（?1kb）组装.将三个片段（左侧侧面，右侧侧翼，右侧侧面）分别扩增并分别克隆到进入载体PDONR TM 221P1-P1-P1-P4，PDONR TM 221 P4R-P3R和PDONR TM 221 P3-P2中.通过PCR确认侧翼克隆，验证序列序列.使用LR Clonase?II Plus创建最终组装构建体，以彼此互相组合三个克隆，目的地向量，PET-DEST-TIGR02.线性化载体可用于转化化学态化的原生质体.同源重组后，Pyrg基因补充一种助体营养突变体A1163菌株.请参阅Invitrogen?网关?技术手册 . NR-19437确认板取向和可行性. 96孔板的每个接种孔含有大约60μl的. Coli 培养（菌株DH10B-T1）在Luria Bertani（LB）肉汤中，含有100μg/ ml氨苄青霉的肉汤，其补充有15％甘油. 克隆板被复制使用生物滤泡? fx机器人. 96孔格式的生产增加了相邻井之间的交叉污染风险.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 Resources仅确认随机挑选的克隆的克隆板取向和可存度. Bei资源不确认或验证贡献者提供的单个克隆身份.
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。
品牌标识
可能感兴趣的内容
2023-07-18
2019-06-24
2024-08-16
2022-04-01
2024-08-16
2021-12-21
2024-03-25
2021-12-21
2022-03-22
2022-04-01
2021-12-21
2021-12-21
2022-04-01
2024-01-21
2021-12-21
2021-12-21