Team:Chiba/jk/γ/week 6

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(Difference between revisions)

Latest revision as of 15:48, 9 October 2008

>実験ログ
>出力班

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	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

>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

@@ Line 255: / Line 255: @@
 <br>->30min 37℃
 <br>Gel cheack
-<br>Vector:BBa_R0062
+<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</td>
+<td>each(digestion)</td><td>I0462(natural)</td>
 </tr>
 <tr>
@@ Line 351: / Line 355: @@
 <br>->3hour 37℃
 <br>->Transformation
 == 26,September,2008 ==