Template:Team:UC Berkeley/Notebook/DV construction

From 2008.igem.org

(Difference between revisions)
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Construction of [<Phns>] Basic Part K112701
Construction of [<Phns>] Basic Part K112701
-
PCR dv001/dv004 on Phns             (150 bp, gp = A)
+
PCR dv001/dv004 on Phns from E. coli K12 MG1655            (150 bp, gp = A)
-
PCR dv003/dv006 on Phns             (225 bp, gp = B)
+
PCR dv003/dv006 on Phns from E. coli K12 MG1655            (225 bp, gp = B)
-
PCR dv005/dv002 on Phns             (342 bp, gp = C)
+
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 mv001/dv006 on A+B              (355 bp, gp = D)
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Mixing of dv007/dv008  (27bp, EcoRI/BamHI/DpnI)
Mixing of dv007/dv008  (27bp, EcoRI/BamHI/DpnI)
-
Sub into (vector?)pBca1256                      (EcoRI/BamHI, 2472+27, L)
+
Sub into pBca1256                      (EcoRI/BamHI, 2472+27, L)
Product is pBca1256-K112702
Product is pBca1256-K112702
-----------------------------------------------
-----------------------------------------------
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==K112703==
==K112703==
<pre>
<pre>
-
Mix dv009/dv010 on pBca9145-Bca1117    (41bp, EcoRI/BamHI/DpnI)
+
Mix dv009/dv010                       (41bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+41, L)
Sub into pBca1256                      (EcoRI/BamHI, 2472+41, L)
Product is pBca1256-K112703
Product is pBca1256-K112703
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==K112704==
==K112704==
<pre>
<pre>
-
Wobble PCR of dv011/dv012 (no template) (60 bp, EcoRI/BamHI/DpnI)
+
Wobble PCR of dv011/dv012 (60 bp, EcoRI/BamHI/DpnI)
Sub into pBca1256 (EcoRI/BamHI)
Sub into pBca1256 (EcoRI/BamHI)
Product is pBca1256-K112704
Product is pBca1256-K112704
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==K112705==
==K112705==
<pre>
<pre>
-
Mix dv013/dv014 (no template)          (47 bp, EcoRI/BamHI/DpnI)
+
Mix dv013/dv014 on pBca1              (47 bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+47, L)
Sub into pBca1256                      (EcoRI/BamHI, 2472+47, L)
Product is pBca1256-K112705
Product is pBca1256-K112705
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==K112706==
==K112706==
<pre>
<pre>
-
PCR of dv015/dv016 (template?)         (500bp, EcoRI/BamHI/DpnI)
+
PCR of dv015/dv016 on pBca1037         (500bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+ 500, L)
Sub into pBca1256                      (EcoRI/BamHI, 2472+ 500, L)
Product is pBca1256-K112706
Product is pBca1256-K112706
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==K112707==
==K112707==
<pre>
<pre>
-
PCR of dv017/dv018 (template?)         (499bp, EcoRI/BamHI/DpnI)
+
PCR of dv017/dv018 on pBca1037         (499bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+499, L)
Sub into pBca1256                      (EcoRI/BamHI, 2472+499, L)
Product is pBca1256-K112707
Product is pBca1256-K112707
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==K112708==
==K112708==
<pre>
<pre>
-
PCR of dv019/dv020 (template?)         (225bp, EcoRI/BamHI/DpnI)
+
PCR of dv019/dv020 on  pBjh1205         (225bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                        (EcoRI/BamHI, 2472+225, L)
Sub into pBca1256                        (EcoRI/BamHI, 2472+225, L)
Product is pBca1256-K112708
Product is pBca1256-K112708
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==K112710==
==K112710==
<pre>
<pre>
-
PCR of dv023/dv024                     (122bp, EcoRI/BamHI/DpnI)
+
PCR of dv023/dv024 Bca1041                  (122bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+122, L)
Sub into pBca1256                      (EcoRI/BamHI, 2472+122, L)
Product is pBca1256-K112710
Product is pBca1256-K112710

Revision as of 01:13, 11 June 2008

Contents


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
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