[問題] 分生問題---設計載體

看板Biotech (生命科學)作者tzuling0815 (豔陽天.晴)時間19年前 (2006/04/05 19:55)推噓1(1推 0噓 0→)

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有一個分生的問題想請教大家... 之前有問過了...但是都沒有人回我><" 以下是原文... 主旨是要我們做一個載體.並希望此在體可以把PCR產品的DNA放入載體中（可克隆）＊問題：該如何將載體只在3'處多一個T... 並找出片段有多長.方向為何？如何讓載體切開後黏不起來... 我找過很多資料還是不懂該怎麼回答問題...請求大家幫我解惑吧！！先謝謝囉！！ 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. -- ※ 發信站: 批踢踢實業坊(ptt.cc) ◆ From: 218.168.136.194