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(μL) 1
Fwd primer(μL) 1.5
Rev primer(μL) 1.5
Taq(DNA pol)(μL) 0.2
dNTPmix(μL) 3
thermo pol Buffer(μL) 3
dH2O(μL) 19.8
TOTAL(μL) 30


->95°C 5 min ->(95°C 1 min 50°C 30sec 72°C 1 min)×25 ->72°C 10 min


Digestion Test
BBa_I6206
BBa_I6101
BBa_I6042
BBa_I0462

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


->30 min 37°C
Gel check
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(μL) 101
Loading Dye(μL) 21
dH2O(μL) -4
TOTAL(μL) 126


Digestion
BBa_I0462

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


->3 hours 37°C
->Zymo ->10μL


Ligation
Vector:BBa_I7100
Insert:BBa_R0062
Insert:BBa_I0462

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


->3 hours 37°C
->Transformation