Team:UT-Tokyo/Sudoku notebook 2

From 2010.igem.org

(Difference between revisions)
(Make construct)
(Make construct)
Line 93: Line 93:
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==== 7/3 ====
+
==== 8/5 ====
 +
 
 +
'''Enzyme digestion'''
 +
 
 +
1234(107.2ng/uL)
'''Gel extract'''
'''Gel extract'''
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2-3, 11 plasmid (7/2 enzyme cut)
+
sudoku (1234) 6.6ng/ul
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Label : 100703 X ゲル抽出
+
'''Ligation'''
 +
0(1234)&iii

Revision as of 02:05, 19 September 2010

Sudoku

Abstract Construct Experiment Notebook Result

Notebook

Pre-experiment

AUGUST!!!!!


AUGUST!!!!!



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



AUGUST!!!!!

Make construct

8/2

Sequencing

sequel to 7/31

add HiDi 20μL

A6 I pre, B6 I sur,C6 II pre

D6 II sur, E6 III pre, F6 III sur


8/4

Miniprep

12-Ⅰ 58.7ng/uL

12-Ⅱ 55.9ng/uL

13ーⅠ 48.7ng/uL

13ーⅡ 48.5ng/uL


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) 


Ligation

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

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


Gel extract

i 1-99234 8.8ng/uL

ii 1-9234 7.1ng/uL

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


8/5

Enzyme digestion

1234(107.2ng/uL)

Gel extract

sudoku (1234) 6.6ng/ul

Ligation

0(1234)&iii


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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