Template:Team:UC Berkeley/Notebook/DV construction

From 2008.igem.org

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==K112702==
==K112702==
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<pre>
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Construction of <His-Tag! Basic Part K112702
Construction of <His-Tag! Basic Part K112702
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dv007  Forward biobricking of <His-Tag!    CATCATCATCATCATCATtaaGGATCC
dv007  Forward biobricking of <His-Tag!    CATCATCATCATCATCATtaaGGATCC
dv008  Reverse biobriquing of <His-Tag!    GGATCCttaATGATGATGATGATGATG
dv008  Reverse biobriquing of <His-Tag!    GGATCCttaATGATGATGATGATGATG
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PCR dv009/dv010 on pBca9145-Bca1117    (1060bp, EcoRI/BamHI/DpnI)
 
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Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
 
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Product is pBca1256-K112100
 
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dv009 Forward Biobricking of His-Tag>
 
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ccataGAATTCatgAGATCTatgCATCATCATCATCATCAT
 
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dv010  Reverse Biobricking of His-Tag>
 
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ATGATGATGATGATGATGcatAGATCTcatGAATTCtatgg
 
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<pre>
 
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PCR bx1/bx2 on Bjh1301 (688 bp, EcoRI/BamHI/DpnI)
 
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Sub into pBca1256 (EcoRI/BamHI)
 
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Product is pBca1256-K112000
 
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bx1 Forward Biobricking of {T4 Holin!}
 
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ccagtGAATTCatgAGATCTatggcagcacctagaatatc
 
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bx2 Reverse Biobricking of {T4 Holin!}
 
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cgttaGGATCCttatttagcccttcctaatattctggccgcc
 
</pre>
</pre>

Revision as of 23:19, 10 June 2008

Contents


K112701

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

K112702

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