August/10 August 2009

From 2009.igem.org

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 CaCl2)


3.digestion and ligation
digestion with restriction enzyme

Vector1Insert1
1-23L122-16H6
EcoRI1EcoRI1
SpeI1XbaI1
No.2 Buffer2No.22
dH2O4dH2O10
total20uLtotal20uL


Vector2Insert2
1-23L123-18O5
EcoRI1EcoRI1
SpeI1XbaI1
No.2 Buffer2No.22
dH2O4dH2O11
total20uLtotal20uL




Osaka090810.jpg

vector1 insert1 vector2 insert2 ladder

failure


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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