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pEASY®-Blunt Zero Cloning Vector contains a suicide gene. Ligation of PCR fragment disrupts the expression of the gene. Cells that contain non-recombinant vector are killed upon plating. Therefore, blue/white selection is not required.
Using the vector only for transformation is not a recommended negative control. The process of TOPO-adaptation is not a 100% process, therefore, there will be “vector only” present in your mix, and colonies will be obtained. Please try the suggestions below to increase the number of colon...
- The molar ratio of vector:insert in the ligation reaction may be incorrect. Estimate the concentration of the PCR product. Set up the ligation reaction with a 1:1 or 1:3 vector:insert molar ratio. On a typical plate there are a few white colonies which do not contain insert. These ...
操作过程省去冰浴、热激和1 h 复苏。 应用 基因克隆 使用说明 请参考产品说明进行实验操作。 组分 pUCI-Blunt Zero cloning Vector (30 ng/μL),1 kb control insert (40 ng/μL),10X Enhancer ,M13F Primer 10 μM,M13R Primer 10 μM,Sterilized ddH2O...
Zero Blunt TOPO PCR Cloning kits for subcloning provide a highly efficient, 5-minute cloning reaction for the direct insertion of blunt-ended PCR products amplified with proofreading thermostable polymerases into a plasmid vector. Each kit
用于平末端PCR产物的快速克隆;克隆后PCR产物的快速测序(使用M13F/M13R引物)。 产品信息 货号 10909ES20 规格 20T 组分信息 组分编号 组分名称 10909ES20 10909-A pESI-Bluntvector (30 ng/μL) 20 μL 10909-B 1 kb control insert (40 ng/μL) ...
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。 克隆后PCR产物的快速测序(使用M13F/M13R引物)。
The pCR-Blunt vector is prepared with blunt ends to accept blunt-ended fragments. Due to the terminal transferase activity of Taq DNA polymerase, PCR products amplified with this enzyme have 3'-A overhangs. In order to clone these products into pCR-Blunt, you would need to polish the ends ...
When cloning a Pfx-amplified PCR product, the insert to vector ratio is an important consideration. The PCR product generally needs to be diluted since Pfx generates a high concentration of product and using too much insert DNA can hamper the TOPO reaction. A 1:1 molar ratio of vector to ...