Team:Chiba/Calendar-Home/25 September 2008

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24 September 2008 <|> 26 September 2008

Laboratory work

Team:Communication

Team:Output


colony PCR
Vector:BBa_R0062 Insert:BBa_E0240
Vector:BBa_R0062

Sample No each
culture 1
Fwd primer 1.5
Rev primer 1.5
Taq(DNA pol) 0.2
dNTPmix 3
thermo pol Buffer 3
dH2O 19.8
TOTAL 30


->95℃ 5min ->(95℃ 1min 50℃ 30sec 72℃ 1min)x25 ->72℃ 10min


Digestion Test
BBa_I6206
BBa_I6101
BBa_I6042
BBa_I0462

Sample No each
DNA 2
EcoRⅠ 0.2
EcoRⅠ Buffer 1
dH2O 7
TOTAL 30


->30min 37℃
Gel cheack
BBa_I6206(digestion product)
BBa_I6101(digestion product)
BBa_I6042(digestion product)
BBa_I0462(digestion product)
BBa_I0642(natural)

Sample No each(digestion)I0462(natural)
DNA 101
Loading Dye 21
dH2O -4
TOTAL 126


Digestion
BBa_I0462

DNA 25
XbaⅠ 1
PstⅠ 1
Buffer 3 4
BSA 4
dH2O 5
TOTAL 40


->3hour 37℃
->Zymo ->10μl


Ligation
Vector:BBa_I7100
Insert:BBa_R0062
Insert:BBa_I0462

I7100 1.31.3
R0062 2-
I0462 3.3-
Ligase 11
Ligase Buffer 22
dH2O 0.45
TOTAL 1010


->3hour 37℃
->Transformation