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- | {{UT-Tokyo_HEADER}}
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- | <div id="main_all">
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- | = '''Sudoku''' =
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- | == '''Data''' ==
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- | == Pre-experiment ==
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- | === Make construct ===
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- | ==== 7/1 ====
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- | '''Transformation'''
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- | 2-3 plasmid(6/30 ligation)
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- | <6/30 transformation → fail>
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- | '''Inoculation'''
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- | 11 plasmid(6/30 thaw, transformation)
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- | '''Enzyme cut'''
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- | 2, 3, 5, 7 plasmid(6/25 miniprep)
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- | 2 : 53.9 ng/uL → about 15 uL(shortage) / H / SP(2 uL)
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- | 3 : 124.2 ng/uL → 8 uL / M / XP(2 uL)
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- | 5 : 81.3 ng/uL → 12.3 uL / H / SP(2 uL)
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- | 7 : 85.4 ng/uL → 11.7 uL / H / SP(2 uL)
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- | '''Gel extract'''
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- | 2, 3, 5, 7 plasmid(7/1 enzyme cut)
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- | Label: 100701 数X(2,3,5,9) ゲル抽出
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- | ==== 7/2 ====
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- | '''Miniprep'''
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- | 2-3, 11 plasmid(7/1 inoculation)
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- | 2-3 → 105 ng/uL
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- | 11(1st) → 191.3 ng/uL
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- | 11(2nd) → 49.5 ng/uL
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- | '''Enzyme cut'''
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- | 2-3, 11 plasmid(7/2 miniprep)
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- | 2-3 : 9.5 uL / H / ES(1.5 uL)
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- | 11(1st) : 5.2 uL / H / ES(1.5 uL) → Judged failure by cut check → miniprep again
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- | 11(2nd) : 20.2 uL / H / ES(1.5 uL)
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- | === Check test ===
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- | Check test
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- | {| border="1"
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- | | Number || Name || Link to BioBrick || Plate coordinate || Vector || Code length
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- | |-
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- | | 1 || T7 promoter ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_I712074 BBa_I712074]
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- | || plate1-6N || pSB1AK8 || 46bp
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- | |-
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- | | 2 || rbs ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0030 BBa_B0030]
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- | || plate1-1H || pSB1A2 || 15bp
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- | |-
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- | | 3 || cre recombinase ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_J61047 BBa_J61047]
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- | || plate1-5D || pSB1A2 || 1037bp
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- | |-
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- | | 4 || double terminator ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014 BBa_B0014]
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- | || plate2-24C || pSB1AK3 || 95bp
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- | |-
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- | | 5 || lox66 recombinase site ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_I718017 BBa_I718017]
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- | || plate1-17J || pSB1A2 || 34bp
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- | |-
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- | | 6 || Kan resistance (rev) ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_J31002 BBa_J31002]
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- | || plate1-2K || pSB1A2 || 816bp
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- | |-
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- | | 7 || rbs (rev) ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014 BBa_B0014]
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- | || plate1-1J || pSB1A2 || 15bp
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- | |-
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- | | 8 || lox71 recombinase site ||
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- | order-made
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- | || --- || --- || 34bp
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- | |-
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- | | 9 || single terminator ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_B1006 BBa_B1006]
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- | || plate1-4H || pSB1AK3 || 39bp
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- | |-
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- | | 10 || constant express promoter ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_J23119 BBa_J23119]
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- | || plate1-18A || pSB1A2 || 35bp
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- | |-
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- | | 11 || Tet resistance (rev) ||
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- | [http://partsregistry.org/wiki/index.php?title=Part:BBa_J31006 BBa_J31006]
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- | || plate1-1N || pSB1A2 || 1191bp
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- | |}
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- | == Main construct ==
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- | === Research of SP6 RNApol cDNA and F plasmid ===
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- | Research of SP6 RNApol cDNA and F plasmid
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- | === Convert lox71(rev) to lox66(rev) ===
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- | Convert lox71(rev) to lox66(rev)
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- | === Make Hin(rev) ===
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- | Make Hin(rev)
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- | === Revise MS2 gene ===
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- | Revise MS2 gene
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- | === Make location(rev)-rbs(rev)-location(rev) ===
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- | Make location(rev)-rbs(rev)-location(rev)
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- | </div>
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- | {{UT-Tokyo_FOOTER}}
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