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规格:10×100 μL
产品简介:
本产品专为高效克隆不稳定DNA片段、毒性基因及甲基化修饰DNA而设计,其独特的基因型使其在克隆富含甲基化修饰的DNA片段时更出色。诱导启动子驱动的改良pcnB基因,确保了质粒的拷贝数可控。[mcrA,A(mrr-hsdRMS-mcrBC)]基因型完整缺失甲基化限制系统,彻底消除对甲基胞嘧啶/甲基腺嘌呤修饰DNA的切割活性,适合于克隆富含甲基胞嘧啶或甲基腺嘌呤的DNA;recA1和endA1的突变有利于插入DNA的稳定和高纯度质粒DNA的提取;lacZ△M15用于重组克隆的蓝白斑筛选实验;tonA赋予其抗噬菌体T1和T5的能力;rpsL赋予其链霉素抗性。本产品经优化的感受态制备工艺制备而成,使用pUC19质粒DNA检测,转化效率高达5×10^⁷cfu/μg。
产品应用:
◆ 本产品适用于各种不稳定DNA或毒性基因的克隆。
产品特点:
◆ 转化效率高达5×10^⁷cfu/μg;
◆ 链霉素抗性;
◆ 可用于蓝白斑筛选。
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文献和实验Preparing chemically competent cells
Materials Plate of cells to be made competent TSS buffer LB media Ice Glassware & Equipment Falcon tubes 500μl Eppendorf tubes, on ice 200ml conical flask 200μl pipetman or repeating
TOP10 chemically competent cells
Detergent is a major inhibitor of competent cell growth and transformation. Glass and plastic must be detergent free for these protocols. The easiest way to do this is to avoid washing glassware, and simply rinse it out. Autoclaving glassware filled
CCMB buffer to make chemical competent cell
TOP10 chemically competent cells ( modified from http://openwetware.org/wiki/TOP10_chemically_competent_cells#CCMB80_buffer) I also use this protocol to make Ecoli compent cell. It is pretty good. You can start from the fresh clone on plate











