有一个分生的问题想请教大家... 之前有问过了...但是都没有人回我><" 以下是原文... 主旨是要我们做一个载体.并希望此在体可以把PCR产品的DNA放入载体中（可克隆） ＊问题：该如何将载体只在3'处多一个T... 并找出片段有多长.方向为何？ 如何让载体切开後黏不起来... 我找过很多资料还是不懂该怎麽回答问题...请求大家帮我解惑吧！！ 先谢谢罗！！ Design a vector from mp18 vector in the reference material to clone a PCR product, this vector can be called your own “TA cloning” vector. And you are going to sell this open vector to earn big money. TA cloning is brought about by the terminal transferase activity of certain type of DNA polymerase such as the Taq polymerase. This enzyme adds a single, 3'-A overhang to each end of the PCR product. As a result, the PCR product can be directly cloned into a linearized cloning vector that have single base 3'-T overhangs on each end. Such vectors are called T- vectors. The PCR product with A overhang, is mixed with this vector in high proportion. The complementary overhangs of a "T" vector and the PCR product hybridize. The result is a recombinant DNA, the recombination being brought about by DNA ligase. Use any restriction enzyme you want in the “MCS” region of mp18. Remove any sequence by restriction enzymes. Add any sequence with machine-synthesized oligonucleotides (write down the exact nucleotides you add and the insertion direction). Use the table of compatible ends between different restriction enzymes. Tell me how you open your vector and keep it open for your customer ready to use. --

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