August/10 August 2009

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1.<B>Min prep</b><br>
1.<B>Min prep</b><br>
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Checked the cell cultures transformed and plated out yesterday (8/9) for colony formation and made an approximate count of the number of colonies.
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Result of Nanodrop concentration check
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  (plate number)-(location on plate) (no. of colonies)
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  (sample No.) (concentration)
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  1-6O   35.2 ng/uL  
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  1-6O           35.2 ng/uL  
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  1-18L 41.8 ng/uL  
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  1-18L           41.8 ng/uL  
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  1-10K 39.1 ng/uL  
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  1-10K           39.1 ng/uL  

Revision as of 10:29, 11 August 2009

1.Min prep
Result of Nanodrop concentration check 

(sample No.) (concentration)
1-6O            35.2 ng/uL 
1-18L           41.8 ng/uL 
1-10K           39.1 ng/uL


2.make competent cells(with Ca)


3.digestion and ligation
digestion with restriction enzyme

Insert 1 uL
2-16H 6
EcoRⅠ 1
XbaⅠ 1
10*Buffer No.2 2
dH2O 10
TOTAL 20


Vector 1 uL
1-23L 12
EcoRⅠ 1
SpeⅠ 1
10*Buffer No.2 2
dH2O 4
TOTAL 20



Insert 2 uL
3-18O 5
EcoRⅠ 1
XbaⅠ 1
10*Buffer No.2 2
dH2O 11
TOTAL 20


Vector 2 uL
1-23L 12
EcoRⅠ 1
SpeⅠ 1
10*Buffer No.2 2
dH2O 4
TOTAL 20

↓
failured 


digestion with restriction enzyme
insert : E+S 
vector : E+x



4.Transformation
to get new plasmid 

1-23L A 
1-18C A 
1-20F K 
1-12H K 
1-6I  A 
2-6P  A 
1-15N A
used '8/10 competent cell
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