Team:Chiba/jk/γ/week 6
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(→27,September,2008) |
(→25,September,2008) |
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== WEEK 6 == | == WEEK 6 == | ||
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+ | == 21,September,2008 == | ||
== 22,September,2008 == | == 22,September,2008 == | ||
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<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257" | + | <td width="257">DNA</td> |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
<td>2</td> | <td>2</td> | ||
</tr> | </tr> | ||
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<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257" | + | <td width="257">DNA</td> |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
<td>1</td> | <td>1</td> | ||
</tr> | </tr> | ||
Line 136: | Line 131: | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>6</td> |
</tr> | </tr> | ||
</table> | </table> | ||
Line 260: | Line 255: | ||
<br>->30min 37℃ | <br>->30min 37℃ | ||
<br>Gel cheack | <br>Gel cheack | ||
- | <br> | + | <br>BBa_I6206(digestion product) |
+ | <br>BBa_I6101(digestion product) | ||
+ | <br>BBa_I6042(digestion product) | ||
+ | <br>BBa_I0462(digestion product) | ||
+ | <br>BBa_I0642(natural) | ||
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
- | <td>each(digestion)</td><td>I0462</td> | + | <td>each(digestion)</td><td>I0462(natural)</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 356: | Line 355: | ||
<br>->3hour 37℃ | <br>->3hour 37℃ | ||
<br>->Transformation | <br>->Transformation | ||
+ | |||
== 26,September,2008 == | == 26,September,2008 == | ||
Latest revision as of 15:48, 9 October 2008
Contents |
WEEK 6
21,September,2008
22,September,2008
Time delay
23,September,2008
Digestion test
BBa_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
Sample No | ① | ② | ③ | ④ | ⑤ |
DNA | 25 | 25 | 25 | 25 | 25 |
EcoRⅠ | - | - | 1 | 1 | - |
SpeⅠ | 1 | - | 1 | 1 | - |
PstⅠ | 1 | 1 | 1 | - | 1 |
XbaⅠ | - | 1 | - | - | 1 | BSA(x10) | 4 | 4 | 4 | - | 4 |
EcoRⅠ Buffer(x10) | - | - | 4 | 4 | - |
Buffer 2 | 4 | - | - | - | - |
Buffer 3 | - | 4 | - | - | 4 |
dH2O | 5 | 5 | 5 | 9 | 5 |
TOTAL | 40 | 40 | 40 | 40 | 40 |
->1hour 37℃
->SAP 1μl(Vector only) ->1hour 37℃ ->30min 65℃
->Zymo ->10μl
Gel cheack
DNA | 1 |
Loading Dye | 1 |
dH2O | 4 |
TOTAL | 6 |
->③&④&⑤のLigationはしない
24,September,2008
Ligation
Vector:BBa_R0062①
Insert:BBa_E0240②
Sample No | ①&② | ① |
DNA(vector) | 1 | 1 |
DNA(insert) | 3.2 | - |
Ligase | 1 | 1 |
Ligase Buffer | 2 | 2 |
dH2O | 2.5 | 6 |
TOTAL | 10 | 10 |
->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
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 | 10 | 1 |
Loading Dye | 2 | 1 |
dH2O | - | 4 |
TOTAL | 12 | 6 |
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 | 1.3 | 1.3 |
R0062 | 2 | - |
I0462 | 3.3 | - |
Ligase | 1 | 1 |
Ligase Buffer | 2 | 2 |
dH2O | 0.4 | 5 |
TOTAL | 10 | 10 |
->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
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