Team:Chiba/jk/γ/week 6

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<br>[[Team:Chiba/jk|>実験ログ]]
+
<br>>[[Team:Chiba/jk|実験ログ]]
-
<br>[[Team:Chiba/jk/γ|>出力班]]
+
<br>>[[Team:Chiba/jk/γ|出力班]]
== WEEK 6 ==
== WEEK 6 ==
 +
 +
 +
== 21,September,2008 ==
== 22,September,2008 ==
== 22,September,2008 ==
Line 16: Line 19:
<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">DNA</td>
-
<td>E0240</td>
+
-
</tr>
+
-
<tr>
+
-
<td>DNA</td>
+
<td>2</td>
<td>2</td>
</tr>
</tr>
Line 48: Line 47:
</tr>
</tr>
</table>
</table>
 +
 +
<br>TRANSFORMATION
 +
<br>BBa_I6107
 +
<br>BBa_I6206
 +
<br>BBa_I6101
 +
<br>BBa_I6042
 +
<br>BBa_I0462
 +
<br>Digestion
<br>Digestion
-
<br>Vector:BBa_E0240
+
<br>Vector:BBa_R0062①
-
<br>Insert:BBa_R0062
+
<br>Insert:BBa_E0240②
 +
<br>Vector:BBa_I7100③
 +
<br>Insert:BBa_R0062④
 +
<br>Insert:BBa_J37033⑤
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No</td>
 +
<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
 +
</tr>
 +
<tr>
 +
<td>DNA</td>
 +
<td>25</td><td>25</td><td>25</td><td>25</td><td>25</td>
 +
</tr>
 +
<tr>
 +
<td>EcoRⅠ</td>
 +
<td>-</td><td>-</td><td>1</td><td>1</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>SpeⅠ</td>
 +
<td>1</td><td>-</td><td>1</td><td>1</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>PstⅠ</td>
 +
<td>1</td><td>1</td><td>1</td><td>-</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>XbaⅠ</td>
 +
<td>-</td><td>1</td><td>-</td><td>-</td><td>1</td>
 +
</tr>
 +
<td>BSA(x10)</td>
 +
<td>4</td><td>4</td><td>4</td><td>-</td><td>4</td>
 +
</tr>
 +
<tr>
 +
<td>EcoRⅠ Buffer(x10)</td>
 +
<td>-</td><td>-</td><td>4</td><td>4</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 2</td>
 +
<td>4</td><td>-</td><td>-</td><td>-</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 3</td>
 +
<td>-</td><td>4</td><td>-</td><td>-</td><td>4</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O</td>
 +
<td>5</td><td>5</td><td>5</td><td>9</td><td>5</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>40</td><td>40</td><td>40</td><td>40</td><td>40</td>
 +
</tr>
 +
</table>
 +
 
 +
<br>->1hour 37℃
 +
<br>->SAP 1μl(Vector only) ->1hour 37℃ ->30min 65℃
 +
<br>->Zymo  ->10μl
 +
<br>Gel cheack
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">DNA</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Loading Dye</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O</td>
 +
<td>4</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>6</td>
 +
</tr>
 +
</table>
 +
 
 +
<br>->③&④&⑤のLigationはしない
 +
 
 +
== 24,September,2008 ==
 +
 
 +
<br>Ligation
 +
<br>Vector:BBa_R0062①
 +
<br>Insert:BBa_E0240②
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No</td>
 +
<td>①&②</td><td>①</td>
 +
</tr>
 +
<tr>
 +
<td>DNA(vector)</td>
 +
<td>1</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>DNA(insert)</td>
 +
<td>3.2</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>Ligase</td>
 +
<td>1</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Ligase Buffer</td>
 +
<td>2</td><td>2</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O</td>
 +
<td>2.5</td><td>6</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>10</td><td>10</td>
 +
</tr>
 +
</table>
 +
 
 +
<br>->TRANSFORMATION
 +
 
 +
 
 +
== 25,September,2008 ==
 +
<br>colony PCR
 +
<br>Vector:BBa_R0062 Insert:BBa_E0240
 +
<br>Vector:BBa_R0062
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No</td>
 +
<td>each</td>
 +
</tr>
 +
<tr>
 +
<td>culture</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Fwd primer</td>
 +
<td>1.5</td>
 +
</tr>
 +
<tr>
 +
<td>Rev primer</td>
 +
<td>1.5</td>
 +
</tr>
 +
<tr>
 +
<td>Taq(DNA pol)</td>
 +
<td>0.2</td>
 +
</tr>
 +
<tr>
 +
<td>dNTPmix</td>
 +
<td>3</td>
 +
</tr>
 +
<tr>
 +
<td>thermo pol Buffer</td>
 +
<td>3</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O</td>
 +
<td>19.8</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>30</td>
 +
</tr>
 +
</table>
 +
 
 +
<br>->95℃ 5min ->(95℃ 1min 50℃ 30sec　72℃ 1min)x25 ->72℃ 10min
 +
 
 +
<br>Digestion Test
 +
<br>BBa_I6206
 +
<br>BBa_I6101
 +
<br>BBa_I6042
 +
<br>BBa_I0462
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No</td>
 +
<td>each</td>
 +
</tr>
 +
<tr>
 +
<td>DNA</td>
 +
<td>2</td>
 +
</tr>
 +
<tr>
 +
<td>EcoRⅠ</td>
 +
<td>0.2</td>
 +
</tr>
 +
<tr>
 +
<td>EcoRⅠ Buffer</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O</td>
 +
<td>7</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>30</td>
 +
</tr>
 +
</table>
 +
 
 +
<br>->30min 37℃
 +
<br>Gel cheack
 +
<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">
 +
<tr>
 +
<td width="257">Sample No</td>
 +
<td>each(digestion)</td><td>I0462(natural)</td>
 +
</tr>
 +
<tr>
 +
<td>DNA</td>
 +
<td>10</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Loading Dye</td>
 +
<td>2</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O</td>
 +
<td>-</td><td>4</td>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>12</td><td>6</td>
 +
</tr>
 +
</table>
 +
 
 +
<br>Digestion
 +
<br>BBa_I0462
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">DNA</td>
 +
<td>25</td>
 +
</tr>
 +
<tr>
 +
<td>XbaⅠ</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>PstⅠ</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 3</td>
 +
<td>4</td>
 +
</tr>
 +
<tr>
 +
<td>BSA</td>
 +
<td>4</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O</td>
 +
<td>5</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>40</td>
 +
</tr>
 +
</table>
 +
 
 +
<br>->3hour 37℃
 +
<br>->Zymo ->10μl
 +
 
 +
<br>Ligation
<br>Vector:BBa_I7100
<br>Vector:BBa_I7100
<br>Insert:BBa_R0062
<br>Insert:BBa_R0062
-
<br>Insert:BBa_J37033
+
<br>Insert:BBa_I0462
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">I7100</td>
 +
<td>1.3</td><td>1.3</td>
 +
</tr>
 +
<tr>
 +
<td>R0062</td>
 +
<td>2</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>I0462</td>
 +
<td>3.3</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>Ligase</td>
 +
<td>1</td><td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Ligase Buffer</td>
 +
<td>2</td><td>2</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O
 +
<td>0.4</td><td>5</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>10</td><td>10</td>
 +
</tr>
 +
</table>
 +
 
 +
<br>->3hour 37℃
 +
<br>->Transformation
 +
 
 +
== 26,September,2008 ==
 +
 
 +
<br>TIME DELAY TEST
 +
 
 +
<br>colony PCR
 +
<br>Ligation product(26,September)x7
 +
<br>Vector:BBa_R0062
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 +
<tr>
 +
<td width="257">Sample No</td>
 +
<td>each</td>
 +
</tr>
 +
<tr>
 +
<td>culture</td>
 +
<td>1</td>
 +
</tr>
 +
<tr>
 +
<td>Fwd primer</td>
 +
<td>1.5</td>
 +
</tr>
 +
<tr>
 +
<td>Rev primer</td>
 +
<td>1.5</td>
 +
</tr>
 +
<tr>
 +
<td>Taq(DNA pol)</td>
 +
<td>0.2</td>
 +
</tr>
 +
<tr>
 +
<td>dNTPmix</td>
 +
<td>3</td>
 +
</tr>
 +
<tr>
 +
<td>thermo pol Buffer</td>
 +
<td>3</td>
 +
</tr>
 +
<tr>
 +
<td>dH2O</td>
 +
<td>19.8</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>30</td>
 +
</tr>
 +
</table>
 +
 
 +
<br>->95℃ 5min ->(95℃ 1min 50℃ 30sec　72℃ 1min)x25 ->72℃ 10min
 +
<br>->Gel cheack
 +
<br>->MINI Prep
 +
 
 +
== 27,September,2008 ==
 +
 
 +
 
 +
 
 +
 
 +
<br>>[[Team:Chiba/jk/γ/week 5|Last Week]]
 +
<br>>[[Team:Chiba/jk/γ/week 7|Next Week]]

Latest revision as of 15:48, 9 October 2008


>実験ログ
>出力班


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

</tr>

Sample No ①②③④⑤
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

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&②①
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
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 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
>Next Week


ホーム メンバー紹介 プロジェクト紹介 Parts Submitted to the Registry モデリング ノート
Retrieved from "http://2008.igem.org/Team:Chiba/jk/%CE%B3/week_6"