SBB10Ntbk-Paulina Tran: Difference between revisions
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fk0002 Reverse BamHI for FokI gctagGGATCCaaaattgatctccccattgttg | fk0002 Reverse BamHI for FokI gctagGGATCCaaaattgatctccccattgttg | ||
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Revision as of 02:44, 9 February 2010
Construction file for Project Parts:
Construction of FokI+ cleavage domain basic part sbb21 PCR fk0001/fk0003 on FokI (505 bp, gp = A) PCR fk0004/fk0002 on FokI (141 bp, gp = B)
PCR fk0001/fk0002 on A+B (620 bp, EcoRI/BamHI) Digest pBjk2741-Bca1144 (EcoRI/BamHI, 2170+910, L) Product is pBjk2741-ssb21 {<FokI+>}
fk0001 Forward EcoRI and BglII for FokI ccaaaGAATTCgtccAGATCTCAGCTGGTTaaatctgaactggaggag fk0004 (F)Base Substitution FokI ccataaaaccaatTgcaatggcgccg fk0003 (R)Base Substitution FokI cggcgccattgcAattggttttatgg fk0002 Reverse BamHI for FokI gctagGGATCCaaaattgatctccccattgttg
EIPCR tn0001/tn0002R on pBjk2741-Bca1144 (2230 bp, BglII)Product is pBca9145-Btn0001 {Tn5 5’TR} ------- tn0001 EIPCR construction of Tn5 part CCATAagatctGTCGACTGTCTCTTATACACATCTCAACCggatcctaaCTCGCTCCTCaggcttc tn0002R Reverse BglII oligo for Tn5 EIPCR CCAATAGATCTcatgaattcCACTTCAG