Team:UT-Tokyo/Sudoku notebook 2

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Sudoku

Abstract Construct Experiment Notebook Result

Notebook

Pre-experiment

AUGUST!!!!!


AUGUST!!!!!



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



AUGUST!!!!!

Make construct

8/4

Miniprep

12-i 58.7ng/uL

12-ii 55.9ng/uL

13-i 48.7ng/uL

13-ii 48.5ng/uL

(8/4 Inoculation)


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

(8/4 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


8/5

Enzyme digestion

1234(107.2ng/uL)


Gel extract

sudoku (1234) 6.6ng/ul


Transformation

i-iii i & iii

ii-iii ii & iii

0(1234) & iii

(8/4 & 8/5)


Ligation

0(1234)&iii


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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