Template:Team:UC Berkeley/Notebook/DV construction

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Construction of [<Phns>] Basic Part K112701
Construction of [<Phns>] Basic Part K112701
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PCR dv001/dv004 on Phns from E. coli K12 MG1655            (150 bp, gp = A)
+
PCR dv001/dv003 on Phns from E. coli K12 MG1655            (150 bp, gp = A)
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PCR dv003/dv006 on Phns from E. coli K12 MG1655            (225 bp, gp = B)
+
PCR dv004/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 dv005/dv002 on Phns from E. coli K12 MG1655            (342 bp, gp = C)
---------------------------------------------------
---------------------------------------------------
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PCR mv001/dv006 on A+B              (355 bp, gp = D)
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PCR dv001/dv006 on A+B              (355 bp, gp = D)
---------------------------------------------------
---------------------------------------------------
PCR dv001/dv002 on C+D              (697bp, EcoRI/BamHI/DpnI)
PCR dv001/dv002 on C+D              (697bp, EcoRI/BamHI/DpnI)
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dv005  Removing the FIRST EcoRI site in [<Phns>] (F)CGCTTAATAGGGgATTCTCGTAAACAC
dv005  Removing the FIRST EcoRI site in [<Phns>] (F)CGCTTAATAGGGgATTCTCGTAAACAC
dv006  Removing the FIRST EcoRI site in [<Phns>] (R)GTGTTTACGAGAATcCCCTATTAAGCG
dv006  Removing the FIRST EcoRI site in [<Phns>] (R)GTGTTTACGAGAATcCCCTATTAAGCG
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Revision as of 23:56, 11 June 2008

Contents


K112701

Construction of [<Phns>] Basic Part K112701

PCR dv001/dv003 on Phns from E. coli K12 MG1655            (150 bp, gp = A)
PCR dv004/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 dv001/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>]      CgATAgaattcATGagatctGAAATATAGCTGTGCCATCAGCC
dv002	Reverse Biobricking of [<Phns>]      cagtcggatccGCACGAAGAGTACGGATG
dv003  Removing the 2nd EcoRI site in [<Phns>] (R)  GCCAGGAATGTAAGGAATTCAAAATTGTTC
dv004  Removing the 2ND EcoRI site in [<Phns>] (F)  GAACAATTTTGAATcCCTTACATTCCTGGC
dv005  Removing the FIRST EcoRI site in [<Phns>] (F)CGCTTAATAGGGgATTCTCGTAAACAC
dv006  Removing the FIRST EcoRI site in [<Phns>] (R)GTGTTTACGAGAATcCCCTATTAAGCG

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
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