Team:UT-Tokyo/Sudoku notebook 2

From 2010.igem.org

(Difference between revisions)
(Make construct)
(Make construct)
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==== 8/4 ====
==== 8/4 ====
-
 
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'''Miniprep'''
 
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12-i 58.7ng/uL
 
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12-ii 55.9ng/uL
 
-
 
-
13-i 48.7ng/uL
 
-
 
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13-ii 48.5ng/uL
 
-
 
-
(8/4 Inoculation)
 
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'''Inoculation'''
'''Inoculation'''
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iii 1,5,11,7,8,4Ⅱ EXcut (7/29 Miniprep) 
iii 1,5,11,7,8,4Ⅱ EXcut (7/29 Miniprep) 
 +
 +
 +
'''Gel extract'''
 +
 +
i 1-99234  8.8ng/uL
 +
 +
ii 1-9234  7.1ng/uL
 +
 +
iii 1-5-11-7-8-4  18.0ng/uL
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-
'''Gel extract'''
+
'''Miniprep'''
-
i 1-99234  8.8ng/uL
+
12-i 58.7ng/uL  
-
ii 1-9234  7.1ng/uL
+
12-ii 55.9ng/uL  
-
iii 1-5-11-7-8-4  18.0ng/uL
+
13-i 48.7ng/uL
 +
 
 +
13-ii 48.5ng/uL  
 +
 
 +
(8/4 Inoculation)
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sudoku (1234) 6.6ng/ul
sudoku (1234) 6.6ng/ul
 +
 +
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'''Ligation'''
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 +
0(1234)&iii
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(8/4 & 8/5)
(8/4 & 8/5)
-
 
-
 
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'''Ligation'''
 
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-
0(1234)&iii
 
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==== 8/9 ====
==== 8/9 ====
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''''''
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''''''
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''''''
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 +
''''''
 +
==== 7/5 ====
==== 7/5 ====
==== 7/5 ====
==== 7/5 ====

Revision as of 02:32, 19 September 2010

Sudoku

Abstract Construct Experiment Notebook Result

Notebook

Pre-experiment

AUGUST!!!!!


AUGUST!!!!!



AUGUST!!!!! AUGUST!!!!!



AUGUST!!!!!

Make construct

8/4

Inoculation

sudoku 12-i & 12-ii

sudoku 13-i & 13-ii

(8/3 Transformation)


Enzyme digestion

i 1-99234Ⅰ EScut (7/24 Miniprep & Sequencing)

ii 1-9234Ⅰ EScut (7/21 Miniprep / 7/24 Sequencing)

iii 1,5,11,7,8,4Ⅱ EXcut (7/29 Miniprep) 


Gel extract

i 1-99234 8.8ng/uL

ii 1-9234 7.1ng/uL

iii 1-5-11-7-8-4 18.0ng/uL


Ligation

i-iii i:6ul iii:6ul ligationmix 12ul

ii-iii ii:6ul iii:4ul ligationmix 10ul

(8/4 Miniprep)


Miniprep

12-i 58.7ng/uL

12-ii 55.9ng/uL

13-i 48.7ng/uL

13-ii 48.5ng/uL

(8/4 Inoculation)


8/5

Enzyme digestion

1234(107.2ng/uL)(8/4 gel extract)


Gel extract

sudoku (1234) 6.6ng/ul


Ligation

0(1234)&iii


Transformation

i-iii i & iii

ii-iii ii & iii

0(1234) & iii

(8/4 & 8/5)


8/6

Inoculation

0-iii,(i-iii),ii-iii (8/5 transformation)


transformation

i-iii (8/4 ligation)


8/9

'

'

'

'

7/5

7/5

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

Main construct

Research of SP6 RNApol cDNA and F plasmid

Convert lox71(rev) to lox66(rev)

Make Hin(rev)

Revise MS2 gene

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

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