Kinase-subcloning-Method
Clone # 200
Digest pFastHacI with
HindIII and SalI
HindIII PstI Xho1 StrepII tag broken Xho1
5' agctt CTG CAG ctc gag TGG AGC CAC CCG CAG TTC GAA AAG TgA a 3'
3' a GAC GTC gag ctc ACC TCG GTG GGC GTC AAG CTT TTC AcT tag ct 5'
StrepII-T: agcttCTGCAGctcgagTGGAGCCACCCGCAGTTCGAAAAGTgAa
StrepII-B: tcgatTcACTTTTCGAACTGCGGGTGGCTCCActcgagCTGCAGa
anneal these two oligos and ligate to HindIII and Xho1 digested pDB.His.GST
digestion with Xba1+ Pst1
--Paul.xu 18:13, 6 May 2011 (EST)