Team:UT-Tokyo/Sudoku data

From 2010.igem.org

(Difference between revisions)
Line 14: Line 14:
'''Transformation'''
'''Transformation'''
-
2-3 plasmid(6/30 ligation)
+
2-3 plasmid (6/30 ligation)
<6/30 transformation → fail>
<6/30 transformation → fail>
Line 21: Line 21:
'''Inoculation'''
'''Inoculation'''
-
11 plasmid(6/30 thaw, transformation)
+
11 plasmid (6/30 thaw, transformation)
'''Enzyme cut'''
'''Enzyme cut'''
-
2, 3, 5, 7 plasmid(6/25 miniprep)
+
2, 3, 5, 7 plasmid (6/25 miniprep)
-
2 : 53.9 ng/uL → about 15 uL(shortage) / H / SP(2 uL)
+
2 : 53.9 ng/uL → about 15 uL (shortage) / H / SP (2 uL)
-
3 : 124.2 ng/uL → 8 uL / M / XP(2 uL)
+
3 : 124.2 ng/uL → 8 uL / M / XP (2 uL)
-
5 : 81.3 ng/uL → 12.3 uL / H / SP(2 uL)
+
5 : 81.3 ng/uL → 12.3 uL / H / SP (2 uL)
-
7 : 85.4 ng/uL → 11.7 uL / H / SP(2 uL)
+
7 : 85.4 ng/uL → 11.7 uL / H / SP (2 uL)
'''Gel extract'''
'''Gel extract'''
-
2, 3, 5, 7 plasmid(7/1 enzyme cut)
+
2, 3, 5, 7 plasmid (7/1 enzyme cut)
-
Label: 100701 数X(2,3,5,9) ゲル抽出
+
Label : 100701 数X (2,3,5,9) ゲル抽出
Line 49: Line 49:
'''Miniprep'''
'''Miniprep'''
-
2-3, 11 plasmid(7/1 inoculation)
+
2-3, 11 plasmid (7/1 inoculation)
2-3 → 105 ng/uL
2-3 → 105 ng/uL
-
11(1st) → 191.3 ng/uL
+
11 (1st) → 191.3 ng/uL
-
11(2nd) → 49.5 ng/uL
+
11 (2nd) → 49.5 ng/uL
'''Enzyme cut'''
'''Enzyme cut'''
-
2-3, 11 plasmid(7/2 miniprep)
+
2-3, 11 plasmid (7/2 miniprep)
-
2-3 : 9.5 uL / H / ES(1.5 uL)
+
2-3 : 9.5 uL / H / ES (1.5 uL)
-
11(1st) : 5.2 uL / H /  ES(1.5 uL) → Judged failure by cut check → miniprep again
+
11 (1st) : 5.2 uL / H /  ES (1.5 uL) → Judged failure by cut check → miniprep again
-
11(2nd) : 20.2 uL / H /  ES(1.5 uL)
+
11 (2nd) : 20.2 uL / H /  ES (1.5 uL)
 +
 +
==== 7/3 ====
 +
 +
'''Gel extract'''
 +
 +
2-3, 11 plasmid (7/2 enzyme cut)
 +
 +
Label : 100703 X ゲル抽出
 +
 +
 +
 +
==== 7/5 ====
 +
 +
'''Ligation'''
 +
 +
2-3 (7/3 enzyme cut) & 4 / 11 (7/3 enzyme cut) & 7
 +
 +
2-3 : 37.5 ng/uL → 5 uL & 4 : 5.2 ng/uL → 5 uL
 +
 +
11 : 4.2 ng/uL → 7.5 uL & 7 : Unknown → 2.5 uL
 +
 +
 +
'''Transformation'''
 +
 +
2-3-4 plasmid (7/5 ligation) / 11-7 plasmid (7/5 ligation)
Line 77: Line 102:
-
{| border="1"
 
-
| Number || Name || Link to BioBrick || Plate coordinate || Vector || Code length
 
-
|-
 
-
| 1 || T7 promoter ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I712074  BBa_I712074]
 
-
|| plate1-6N || pSB1AK8 || 46bp
 
-
|-
 
-
| 2 || rbs ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0030  BBa_B0030]
 
-
|| plate1-1H || pSB1A2 || 15bp
 
-
|-
 
-
| 3 || cre recombinase ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J61047  BBa_J61047]
 
-
|| plate1-5D || pSB1A2 || 1037bp
 
-
|-
 
-
| 4 || double terminator ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014  BBa_B0014]
 
-
|| plate2-24C || pSB1AK3 || 95bp
 
-
|-
 
-
| 5 || lox66 recombinase site ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I718017  BBa_I718017]
 
-
|| plate1-17J || pSB1A2 || 34bp
 
-
|-
 
-
| 6 || Kan resistance (rev) ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J31002  BBa_J31002]
 
-
|| plate1-2K || pSB1A2 || 816bp
 
-
|-
 
-
| 7 || rbs (rev) ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B0014  BBa_B0014]
 
-
|| plate1-1J || pSB1A2 || 15bp
 
-
|-
 
-
| 8 || lox71 recombinase site ||
 
-
order-made
 
-
|| --- || --- || 34bp
 
-
|-
 
-
| 9 || single terminator ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_B1006  BBa_B1006]
 
-
|| plate1-4H || pSB1AK3 || 39bp
 
-
|-
 
-
| 10 || constant express promoter ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J23119  BBa_J23119]
 
-
|| plate1-18A || pSB1A2 || 35bp
 
-
|-
 
-
| 11 || Tet resistance (rev) ||
 
-
[http://partsregistry.org/wiki/index.php?title=Part:BBa_J31006  BBa_J31006]
 
-
|| plate1-1N || pSB1A2 || 1191bp
 
-
|}
 

Revision as of 14:58, 23 August 2010

Sudoku

Data

Pre-experiment

Make construct

7/1

Transformation

2-3 plasmid (6/30 ligation)

<6/30 transformation → fail>


Inoculation

11 plasmid (6/30 thaw, transformation)


Enzyme cut

2, 3, 5, 7 plasmid (6/25 miniprep)

2 : 53.9 ng/uL → about 15 uL (shortage) / H / SP (2 uL)

3 : 124.2 ng/uL → 8 uL / M / XP (2 uL)

5 : 81.3 ng/uL → 12.3 uL / H / SP (2 uL)

7 : 85.4 ng/uL → 11.7 uL / H / SP (2 uL)


Gel extract

2, 3, 5, 7 plasmid (7/1 enzyme cut)

Label : 100701 数X (2,3,5,9) ゲル抽出


7/2

Miniprep

2-3, 11 plasmid (7/1 inoculation)

2-3 → 105 ng/uL

11 (1st) → 191.3 ng/uL

11 (2nd) → 49.5 ng/uL


Enzyme cut

2-3, 11 plasmid (7/2 miniprep)

2-3 : 9.5 uL / H / ES (1.5 uL)

11 (1st) : 5.2 uL / H / ES (1.5 uL) → Judged failure by cut check → miniprep again

11 (2nd) : 20.2 uL / H / ES (1.5 uL)


7/3

Gel extract

2-3, 11 plasmid (7/2 enzyme cut)

Label : 100703 X ゲル抽出


7/5

Ligation

2-3 (7/3 enzyme cut) & 4 / 11 (7/3 enzyme cut) & 7

2-3 : 37.5 ng/uL → 5 uL & 4 : 5.2 ng/uL → 5 uL

11 : 4.2 ng/uL → 7.5 uL & 7 : Unknown → 2.5 uL


Transformation

2-3-4 plasmid (7/5 ligation) / 11-7 plasmid (7/5 ligation)


Check test

Check test



Main construct

Research of SP6 RNApol cDNA and F plasmid

Research of SP6 RNApol cDNA and F plasmid

Convert lox71(rev) to lox66(rev)

Convert lox71(rev) to lox66(rev)

Make Hin(rev)

Make Hin(rev)

Revise MS2 gene

Revise MS2 gene

Make location(rev)-rbs(rev)-location(rev)

Make location(rev)-rbs(rev)-location(rev)


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