August/6 October 2009

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1. coloony check

sample no.    no. o f colony
  59                        0
  61                        0
  59                        1
  61                        4
  64                       ++
  65                        6
  X5                        10
  X6                        +
  X7                        10
  X8                        10
 1-24E                    50



2. min prep

sample      conc.
  58            174 ng/uL
  60            220
  62            267
  63            237 
  64            151
  65            75
  67            252
  68            233
  X3           236
  X4           193


3.transformation

sample no. 
11 , 20 , 21 , 2-6O , 1-15J , 1-15L

4.digation
K204052

VectorInsert
1-6I10336
SpeI1XbaI1
PstI1PstI1
No.2 2No.22
dH2O6dH2O6
total20uLtotal20uL


K204067

VectorInsert
1-23L62-16L10
SpeI1XbaI1
PstI1PstI1
No.2 2No.22
dH2O10dH2O6
total20uLtotal20uL

K204067

VectorInsert
1-23L62-16L10
EcoRI1EcoRI1
XbaI1SpeI1
No.22No.22
dH2O10dH2O6
total20uLtotal20uL


K204068

VectorInsert
1-23L101-19B5
EcoRI1EcoRI1
XbaI1SpeI1
No.22No.22
dH2O6dH2O11
total20uLtotal20uL



37°C, 2hr
gel cut & purification
ligation O/N



5.prepared LA pate


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