Template:Team:UC Berkeley/Notebook/DV construction

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Construction of [<Phns>] Basic Part K112701

PCR dv001/dv004 on Phns (150 bp, gp = A) PCR dv003/dv006 on Phns (225 bp, gp = B) PCR dv005/dv002 on Phns (342 bp, gp = C)


PCR mv001/dv006 on A+B (355 bp, gp = D)


PCR dv001/dv002 on C+D (697bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+697) Product is pBca1256-K112701


dv001 Forward Biobricking of [<Phns>] CCATAgaattcATGagatctGAAATATAGCTGTGCCATCAGCC dv002 Reverse Biobricking of [<Phns>] CCATAggatccGCACGAAGAGTACGGATG

dv003 Removing the 1st EcoRI site in [<Phns>] (F) GCCAGGAATGTAAGGgATTCAAAATTGTTC dv004 Removing the 1st EcoRI site in [<Phns>] (R) GAACAATTTTGAATcCCTTACATTCCTGGC dv005 Removing the 2nd EcoRI site in [<Phns>] (F) gctaaagaGttcgccgaagtagc dv006 Removing the 2nd EcoRI site in [<Phns>] (R) gctacttcggcgaaCtctttagc


Construction of <His-Tag! Basic Part K112702

Mixing of dv007/dv008 (27bp, EcoRI/BamHI/DpnI) Sub into (vector?)pBca1256 (EcoRI/BamHI, 2472+27, L) Product is pBca1256-K112110


dv007 Forward biobricking of <His-Tag! CATCATCATCATCATCATtaaGGATCC dv008 Reverse biobriquing of <His-Tag! GGATCCttaATGATGATGATGATGATG


PCR dv009/dv010 on pBca9145-Bca1117 (1060bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+910, L) Product is pBca1256-K112100


dv009 Forward Biobricking of His-Tag> ccataGAATTCatgAGATCTatgCATCATCATCATCATCAT dv010 Reverse Biobricking of His-Tag> ATGATGATGATGATGATGcatAGATCTcatGAATTCtatgg


Contents


K112000

PCR bx1/bx2 on Bjh1301		(688 bp, EcoRI/BamHI/DpnI)
Sub into pBca1256			(EcoRI/BamHI)
Product is pBca1256-K112000
----------------------------------------
bx1		Forward Biobricking of {T4 Holin!}
ccagtGAATTCatgAGATCTatggcagcacctagaatatc
bx2		Reverse Biobricking of {T4 Holin!}
cgttaGGATCCttatttagcccttcctaatattctggccgcc