pTOPO-TA/Blunt Vector(30ng/μL)40μL160μL 1000bp Control(30ng/μL)5μL5μL 10×Enhancer20μL80μL 连接反应的准备: PCR引物使用正常设计的引物即可,不需做任何改变(不能用磷酸化引物)。任意PCR产物(兼容A末端/平末端)均可以直接连接。PCR产物一般建议胶回收纯化(货号:ALH096),这样可以避免后续可能...
Tiosbio®Blunt Zero Vector(20 ng/μL)10μL M13F(-47) (10μM)100μL M13R(-48) (10μM)100μL 无菌水1mL 注意事项 常见问题 问题1:平板克隆数少或不长斑 原因:通常是由于感受态效价较低造成,更换感受态可改善或解决该问题。含有卡那霉素的平板通常长斑数少于含有氨苄青霉素抗生素的平板,平板制作时...
The DNAcan then be ligated directly into the pGEM(R)-T Vector according to standard protocols (7). Following this procedure we obtained greater than 80% recombinants based on blue/white screening and restriction digestion. 展开 被引量: 2 ...
目的基因PCR产物为20ul，我取了5ul电泳检测，条带正确 然后取PCR产物3ul和vector1ul混合（我怀疑就是...
pEASY- Blunt Cloning Vector 1 μl 轻轻混合，室温 (20℃-37℃) 反应5分钟。反应结束后，将离心管...
Traditionally, efficient cloning of blunt-ended fragments into a vector required several steps. The vector needed to be dephosphorylated with eitherCalf Intestinal Alkaline Phosphatase (CIAP). Dephosphorylating the vector keeps it from self-ligating; it also means the vector needs to be pu...
操作过程省去冰浴、热激和1 h复苏。4)可连接长达5 kb目的产物;5)Hieff Clone® Topo-blunt simple vector不含多克隆酶切位点(MCS),使用者在引入后续酶切位点进行酶切操作时,不会受到MCS位点的影响,从而增加酶切效率以及克隆成功率。 产品用途 平端PCR产物的克隆。
Zero Blunt® TOPO® PCR Cloning kits contain pCR™-Blunt II-TOPO vector for use with proofreading enzyme and ccdB gene positive selection. 技术参数 实验方法和说明书下载 Thermo Fisher 测序用 Zero Blunt™ TOPO™ PCR 克隆试剂盒,含 One Shot™ TOP10 Electrocomp™ 大肠杆菌 ...
组成成分 - 4X CloneSmart Vector Premix Includes Buffer, ATP, and one type of ligation-ready Vector - CloneSmart DNA Ligase (2 U/µL) - Positive Control Insert DNA (500 ng/µL lambda HincII) - CloneSmart Sequencing Primers (200 reactions each) SL1 Primer (3.2 pmol/µL) SR2 Primer...
Downstream manipulation (e.g, mutagenesis, subsequent insert addition) is facilitated by the minimal vector size (1.7-2.0 kb). 优点: ▪ Convenient, pre-processed vectors eliminate the need for digestion, gel purification, and dephosphorylation; ▪ Transcription terminators flanking cloning site ...