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运输
wet ice
储存温度
−20°C
一般描述
TargeTron基因敲除系统可通过插入II型内含子为细菌基因的快速特异性破坏提供优化的试剂和实验流程。该方法利用II型内含子的逆转录能力以及一个简单的PCR步骤来重新靶向TargeTron II型内含子，以实现特异性的插入宿主基因组。该技术的实用性已被证明可用于原核基因工程、系统生物学和功能基因组学方法。
与现有的敲除方法相比，TargeTron系统非常快速、简单且强大。该过程包括将目标基因序列加载到Sigma网站（www.sigma-aldrich.com/targetronaccess）上经过充分验证的TargeTron算法、使用一个或多个输出的引物组通过PCR来突变TargeTron内含子模板、将片段连接至pACD4K-C线性载体中、转化宿主、诱导基因破坏、通过卡那霉素抗性选择（插入活化标记物）以及筛选所需的敲除结果。
TargeTron基因敲除系统已在广泛的细菌菌株中得到验证，如大肠杆菌、金黄色葡萄球菌、产气荚膜梭菌、福氏志贺菌、鼠伤寒沙门氏菌和乳酸乳球菌。该系统可经修饰而用于其他物种。
每款TargeTron试剂盒都为指定数量的设计（3 EA或10 EA）提供了访问卡和密码。该密码可使用户能够访问经过广泛验证的TargeTron算法以预测最佳的内含子插入位点，并设计用于突变内含子以插入这些位点的引物。在选择一个或多个所需插入点后，可订购所需的寡核苷酸并使用该试剂盒开始实验过程。3 EA试剂盒中的试剂足以进行3次设计及12次反应。10 EA试剂盒中的试剂足以进行10次设计及40次反应。
应用
特点和优势
简易、简化的实验流程；最多3天内完成敲除
最小的筛选以分离突变体
>90%成功靶向插入
无细胞偶联或特异性宿主因子需要
组分
TargeTron访问卡（在线目标位点选择密码）
内含子PCR模板
EBS通用引物
LacZ对照引物
pACD4K-C线性载体(20 ng/μl)
JumpStart™ REDTaq® ReadyMix®
HindIII 20 U/μl
BsrGI 10 U/μl
10x 限制性酶缓冲液
其他说明
法律信息
推荐
储存分类代码
10 - Combustible liquids
闪点(°F)
Not applicable
闪点(°C)
Not applicable
法规信息
How stable is the knockout using TargeTron, Product TA0100 after several generations?
Data in Lactococcus lactis mutants passaged for 80 generations illustrated that 100% of the mutants were stable.
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products. Once you log onto our website, you will find the price and availability displayed on the product detail page. You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 or view local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
How can I determine possible targets with the Targetron® algorithim? How do I get my primer designs?
Please click here to submit your genomic sequence of interest to Technical Service. We will let you know your available target regions according to the algorithim via email reply.If you are ready to obtain your primer designs, please provide your genomic sequence and your access code from your Targetron® kit in the email and we will provide your designs to you via email reply.
My question is not addressed here, how can I contact Technical Service for assistance?
Ask a Scientist here.
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