Template:Team:UC Berkeley/Notebook/DV construction
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K112701
Construction of [<Phns>] Basic Part K112701 PCR dv001/dv004 on Phns from E. coli K12 MG1655 (150 bp, gp = A) PCR dv003/dv006 on Phns from E. coli K12 MG1655 (225 bp, gp = B) PCR dv005/dv002 on Phns from E. coli K12 MG1655 (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
K112702
Construction of <His-Tag! Basic Part K112702 Mixing of dv007/dv008 (27bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+27, L) Product is pBca1256-K112702 ----------------------------------------------- dv007 Forward biobricking of <His-Tag! CATCATCATCATCATCATtaaGGATCC dv008 Reverse biobriquing of <His-Tag! GGATCCttaATGATGATGATGATGATG
K112703
Mix dv009/dv010 (41bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+41, L) Product is pBca1256-K112703 ----------------------------------------------- dv009 Forward Biobricking of His-Tag> ccataGAATTCatgAGATCTatgCATCATCATCATCATCAT (41 bp) dv010 Reverse Biobricking of His-Tag> ATGATGATGATGATGATGcatAGATCTcatGAATTCtatgg (41 bp)
K112704
Wobble PCR of dv011/dv012 (60 bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI) Product is pBca1256-K112704 ---------------------------------------- dv011 Forward Biobricking of <S-Tag! GATCTAAAGAAACCGCTGCTGCTAAATTCGAACGCCAGCA dv012 Reverse Biobricking of <S-Tag! GGATCCttaCGCTGTCCATGTGCTGGCGTTCGAATTTAGC
K112705
Mix dv013/dv014 on pBca1 (47 bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+47, L) Product is pBca1256-K112705 ----------------------------------------------- dv013 Forward Biobricking of S-Tag> CCATAgaattcATGAGATCTatgGATCTAAAGAAACCGCTGCTGCTA (47 bp) dv014 Reverse Biobricking of S-Tag> CGCTGTCCATGTGCTGGCGTTCGAATTTAGCAGCAGCGGTTTCTTTA (47 bp)
K112706
PCR of dv015/dv016 on pBca1037 (500bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+ 500, L) Product is pBca1256-K112706 ----------------------------------------------- dv015 Forward biobricking of <Pspv2> CCATAgaattcATGAGATCTctagaccttatgcagcgtaaggg dv016 Reverse biobricking of <Pspv2> GGATCCtGATAATGTtTGCAGGGGAATTATTTTGTATTGCC
K112707
PCR of dv017/dv018 on pBca1037 (499bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+499, L) Product is pBca1256-K112707 ----------------------------------------------- dv017 Forward biobricking of <Pspv> CACTGgaattcatgAGATCTccttatgcagcgtaagggccgcaac dv018 Reverse biobricking of <Pspv> CGTACggatccGATAATGTtTGCAGGGGAATT
K112708
PCR of dv019/dv020 on pBjh1205 (225bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+225, L) Product is pBca1256-K112708 ----------------------------------------------- dv019 Forward biobricking of <PfhuA> GGACTgaattcatgAGATCTgctaagcgtgaaataccggatg dv020 Reverse biobricking of <PfhuA> ccgctGGATCCactctgatgtaaagtgaatg
K112709
Wobble PCR of dv021/dv022 (66bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+66, L) Product is pBca1256-K112709 ----------------------------------------------- dv021 Forward biobricking of b1006 ccataGAATTCatgAGATCTaaaaaaaaaccccgcccctgaca dv022 Reverse biobricking of b1006 GGATCCtaaaaaaaaccccgccctgtcaggggcggggtttttt
K112710
PCR of dv023/dv024 Bca1041 (122bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+122, L) Product is pBca1256-K112710 ----------------------------------------------- dv023 Forward biobricking of Bca1041 ccataGAATTCatgAGATCTgCcggcttatcGgtcag dv024 Reverse biobricking of Bca1041 GGATCCtatttcttttgggtatagcg