你的題目...怎麼甚麼條件都沒有啊....也沒說是要做DNA library還是site mutation cloning...還要說仔細一點...faint....
okie..
看你要用甚麼方法去進行clone了. 可以用PCR(in vitro)或者是cell based (in vivo).如果要施行很精密的clone,DNA的size也很重要.
single-stranded DNA如果小過23Kb
1. construct a vector. 可以用phagemid vector或者是M13 vector. 一般vector需要有以下幾個特徵a)under strong promotor; b) have an indicater; c) have a Multiple cloning site (MCS)
2. constuct了vector以後要進行trasfection.Indicater可以指出那一個colony是success transfected.
3. Electrophoresis. 檢查success colony是不是含有Target DNA.
