Team:Tokyo Metropolitan/Notebook/S21

From 2011.igem.org

(Difference between revisions)
(-Targeting-)
(-Visualization-)
 
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-Pre-culture for oriTr-GFP-oriTr
-Pre-culture for oriTr-GFP-oriTr
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 +
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-Colony PCR
 +
 +
Samples which transformed one day ago
 +
 +
 +
-Digestion
 +
 +
Promoter: Spe and Pst
 +
 +
Suppressed expression system: Xba and Pst
 +
 +
Over night

Latest revision as of 13:36, 5 October 2011

-Targeting-

-Plasmid extraction and DNA refining

Extracted plasmid and reined DNA of J23100, P0140 and John-K-Kennedy precultured one day ago.

Results (ng/ul)

J21300 54

P0140 55

John-K-Kennedy

I. 47

II. 46


-Digestion

Digested with P and S for J23100 and John-F-Kennedy, X and P for P0140. Then incubated at 37 deg C.


-Electrophoresis

Checked for treat restriction enzyme

Results

Succeeded to cut J23100, John-F-Kennedy and P0140.


-Gel extraction

Extracted from gel for J23100 and P0140.

Results (ng/ul)

J23100 5

P0140 4


-Ligation and Transformation

Ligated J23100 and P0140 then transformed. Incubated at 37 deg C.


-Visualization-

-Colony PCR for pSB1C3+oriTr-GFP

Did colony PCR for five colony transformed on 20th Sep.


-Electrophoresis for oriTr4,oriTr-GFP 0A,oriTr-GFP 0B

Used plasmid solution, but bands showed thin. Thus restarted from plasmid extraction.


-Pre-culture for oriTr,oriTr-GFP


-Pre-culture for oriTr-GFP-oriTr



-Colony PCR

Samples which transformed one day ago


-Digestion

Promoter: Spe and Pst

Suppressed expression system: Xba and Pst

Over night