Team:Chiba/jk/γ/week 6

From 2008.igem.org

(Difference between revisions)
Line 97: Line 97:
<tr>
<tr>
<td>Buffer 2</td>
<td>Buffer 2</td>
-
<td>4</td>-</td><td>-</td><td>-</td><td>-</td>
+
<td>4</td><td>-</td><td>-</td><td>-</td><td>-</td>
</tr>
</tr>
<tr>
<tr>
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<tr>
<td>Taq(DNA pol)</td>
<td>Taq(DNA pol)</td>
-
<td>o.2</td>
+
<td>0.2</td>
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</tr>
<tr>
<tr>
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<td>thermo pol Buffer</td>
<td>thermo pol Buffer</td>
<td>3</td>
<td>3</td>
 +
</tr>
 +
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<td>dH2O</td>
<td>dH2O</td>
<td>19.8</td>
<td>19.8</td>
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<tr>
<td>TOTAL</td>
<td>TOTAL</td>
-
<td>12</td>
+
<td>12</td><td>6</td>
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</tr>
</table>
</table>

Revision as of 09:21, 28 September 2008


>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


26,September,2008


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






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