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文献和实验利用RACE [Rapid Amplification of cDNA ENDs,cDNA末端的快速扩增],可以很简单地得到mRNA的5'末端序列,然而5'RACE是一个很复杂的技术,它的成功依赖每一步反应的有效完成。cDNA第一链的合成是由一个反义的基因特异性引物(Gene-specific primer,GSP)来起始的,cDNA第一链纯化后,利用末端脱氧核苷酸转移酶(Terminal deoxynucleotidyl transferase,TdT)在cDNA的5'末端加上一个合成
Expression of a cDNA in E.coli in order to prepare an antiserum
an antiserum. As an alternative, the cDNA can be expressed in E.coli as a translational fusion and the expressed protein used to prepare the antiserum. This is normally done as fusion constructs with e.g. -galactosidase ( LacZ
・ cDNA Synthesis (Crawford Lab) mRNA can be converted into DNA (copy DNA, cDNA) by annealing oligo-dT to the 3' poly-A tail that occurs on all eukaryotic mRNA. After the dTs bind to the As, we will use the enzyme "Reverse
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