Team:Chiba/jk/γ/week 6

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>実験ログ
>出力班


Contents

WEEK 6

22,September,2008


Time delay


23,September,2008


Digestion test
BBa_E0240

Sample No E0240
DNA 2
EcoRⅠ 0.5
PstⅠ 0.5
BSA(x10) 1
EcoRⅠ Buffer(x10) 1
dH2O 5
TOTAL 10


TRANSFORMATION
BBa_I6107
BBa_I6206
BBa_I6101
BBa_I6042
BBa_I0462



Digestion
Vector:BBa_R0062①
Insert:BBa_E0240②
Vector:BBa_I7100③
Insert:BBa_R0062④
Insert:BBa_J37033⑤

</tr>

Sample No
DNA 2525252525
EcoRⅠ --11-
SpeⅠ 1-11-
PstⅠ 111-1
XbaⅠ -1--1
BSA(x10) 444-4
EcoRⅠ Buffer(x10) --44-
Buffer 2 4----
Buffer 3 -4--4
dH2O 55595
TOTAL 4040404040


->1hour 37℃
->SAP 1μl(Vector only) ->1hour 37℃ ->30min 65℃
->Zymo ->10μl
Gel cheack

Sample No each
DNA 1
Loading Dye 1
dH2O 4
TOTAL 10


->③&④&⑤のLigationはしない

24,September,2008


Ligation
Vector:BBa_R0062①
Insert:BBa_E0240②

Sample No ①&②
DNA(vector) 11
DNA(insert) 3.2-
Ligase 11
Ligase Buffer 22
dH2O 2.56
TOTAL 1010


->TRANSFORMATION


25,September,2008


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
Vector:BBa_R0062

Sample No each(digestion)I0462
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

26,September,2008


TIME DELAY TEST


colony PCR
Ligation product(26,September)x7
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
->Gel cheack
->MINI Prep

27,September,2008

>Last Week


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