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

File:C6-E3 with HindIII SalI

digestion with Xba1+ Pst1
--Paul.xu 18:13, 6 May 2011 (EST)