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:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] Insert:[http://partsregistry.org/Part:BBa_E0240 BBa_E0240]
Vector:[http://partsregistry.org/Part:BBa_R0062 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
[http://partsregistry.org/Part:BBa_I6206 BBa_I6206]
[http://partsregistry.org/Part:BBa_I6101 BBa_I6101]
[http://partsregistry.org/Part:BBa_I6042 BBa_I6042]
[http://partsregistry.org/Part:BBa_I0462 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
[http://partsregistry.org/Part:BBa_I6206 BBa_I6206](digestion product)
[http://partsregistry.org/Part:BBa_I6101 BBa_I6101](digestion product)
[http://partsregistry.org/Part:BBa_I6042 BBa_I6042](digestion product)
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462](digestion product)
[http://partsregistry.org/Part:BBa_I0642 BBa_I0642](natural)

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


Digestion
[http://partsregistry.org/Part:BBa_I0462 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:[http://partsregistry.org/Part:BBa_I7100 BBa_I7100]
Insert:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]
Insert:[http://partsregistry.org/Part:BBa_I0462 BBa_I0462]

I7100 1.31.3
[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] 2-
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462] 3.3-
Ligase(μL) 11
Ligase Buffer(μL) 22
dH2O(μL) 0.45
TOTAL(μL) 1010


->3 hours 37°C
->Transformation