Team:Chiba/jk/γ/week 2

From 2008.igem.org

(Difference between revisions)

Revision as of 05:38, 24 October 2008

>実験ログ
 >出力班

WEEK 2

24,August 2008

Transformation

[http://partsregistry.org/Part:BBa_J63001 BBa_J63001](yeast optimized YFP)-->colony

Digestion

[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1)①
[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2)②
Menbren Venus YFP③

Sample No.	①	②	③
DNA tamplate	3	3	3
Buffer3	1	1	1
dH2O	6	6	6
EcoRⅠ	0.5	0.5	0.5
TOTAL	10	10	10

-->37℃ 30min

Sample No.	①(afetr digestion)	②(after digestion)	③(afetr digestion)
DNA tamplate	10	10	10
loading Dye	2	2	2
TOTAL	12	12	12

Sample No.	①	②	③
DNA tamplate	2	2	2
loading Dye	1	1	1
dH2O	3	3	3
TOTAL	6	6	6

-->135V 30min

25,August,2008

PCR

[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]①
[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②
BL21③
[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]8④
[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤

>

Sample No	①	②	③	④	⑤
DNA tamplate	1	1	1	1	1
FW primer	5(Venus_YFP_fwd)	5(LacZ_fwd)	5(LacZ_fwd)	5(rLUC_fwd)	5(fLuc_fwd)
VR primer	5(VR)	5(LacZ_rev)	5(LacZ_rev)	5(VR)	5(VR)
dNTPmix	10	10	10	10	10
Thermo pol buffer	10	10	10	10	10
DNA pol(VENT)	1	1	1	1	1
dH2O	68	68	68	68	68
TOTAL	100	100	100	100	100

-->Gel Check

>

Sample No.	①	②	③	④	⑤
DNA tamplate	1	1	1	1	1
loading Dye	1	1	1	1	1
dH2O	4	4	4	4	4
TOTAL	6	6	6	6	6

Mini prep

Venus
[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]

Transformation

Venus
PUC19
pGFPUV
pGEX Venus YFP
Membren Venus YFP

PCR

[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]①

Sample No	①
DNA tamplate	1
FW primer(Venus)	5
VR primer	5
dNTPmix	10
Thermo pol buffer	10
DNA pol(VENT)	1
dH2O	68
TOTAL	100

26,August,2008

Gel Check

[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]

>

Sample No	①	②	③	④	⑤
DNA tamplate	1	1	1	1	1
Loading Dye	1	1	1	1	1
dH2O	4	4	4	4	4
TOTAL	6	6	6	6	6

Digestion
vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]

Sample No	[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
DNA tamplate	100
SpeⅠ	2
PstⅠ	2
BSA(x10)	13
Buffer2	13
TOTAL	130

insert:BBa_J52008,BBa_E2030

Sample No	J52008	E2030
DNA tamplate	30	30
XbaⅠ	1	1
PstⅠ	1	1
DpnⅠ	1	1
Buffer3	4	4
BSA(x10)	4	4
TOTAL	40	40

->37℃　3hour
->SAP
->37℃ 30min
->65℃ 15min
->Zymo Clean
Ligation
vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:BBa_J52008①
vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:BBa_E2030②
Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]③

Sample No	①	②	③
vector	5	5	5
inser	10	7.5	0
Ligase Buffer	4	4	4
Ligase	1	1	1
dH2O	0	2.5	10
TOTAL	20	20	20

-->RT 2hour

-->Transformation(XL10G)

-->37℃ over night

-->colony PCR

-->MINI PREP

28,August,2008

Time Responce(liquid)

29,August,2008

TIME RESPONCE(solid)

30,August,2008

Transformation
mcherry
PCR
BBa_J52008(rLuc)

Sample No	BBa_J52008
DNA tamplate	1
rLuc_fwd	2.5
Rev primer	2.5
Thermo pol Buffer	5
dNTPmix	5
Vent pol	0.5
dH2O	34
TOTAL	50

>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 5: / Line 5: @@
 == 24,August 2008 ==
+[[Team:Chiba/protocol/transformation|Transformation]]
+*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001](yeast optimized YFP)-->colony
+[[Team:Chiba/protocol/digestion|Digestion]]
+*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1)①
+*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2)②
+*Menbren Venus YFP③
-<br>TRANSFORMATION
-<br>BBa_J63001(yeast optimized YFP)-->colony
-<br>Digestion
-<br>BBa_F2620(1)①
-<br>BBa_F2620(2)②
-<br>Menbren Venus YFP③
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample No.</td>
 <td>①</td><td>②</td><td>③</td>
 </tr>
@@ Line 38: / Line 39: @@
 </tr>
 </table>
-<br>->37℃ 30min
+-->37℃ 30min
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample No.</td>
 <td>①(afetr digestion)</td><td>②(after digestion)</td><td>③(afetr digestion)</td>
 </tr>
@@ Line 57: / Line 62: @@
 </tr>
 </table>
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample No.</td>
 <td>①</td><td>②</td><td>③</td>
 </tr>
@@ Line 79: / Line 86: @@
 </tr>
 </table>
-<br>->135V 30min
+-->135V 30min
 == 25,August,2008 ==
-<br>PCR
+[[Team:Chiba/protocol/PCR|PCR]]
-<br>BBa_E2030①
+*[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]①
-<br>BBa_J33202②
+*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②
-<br>BL21③
+*BL21③
-<br>BBa_J52008④
+*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]8④
-<br>BBa_I712019⑤
+*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
@@ Line 128: / Line 137: @@
 </tr>
 </table>
-<br>->Gel
+-->[[Team:Chiba/protocol/gelcheck|Gel Check]]
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample No.</td>
 <td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
 </tr>
@@ Line 152: / Line 164: @@
 </table>
-<br>MINI prep
+[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
-<br>Venus
+*Venus
-<br>BBa_F2620
+*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
-<br>TRANSFORMATRION
-<br>Venus
-<br>PUC19
+[[Team:Chiba/protocol/transformation|Transformation]]
-<br>pGFPUV
+*Venus
-<br>pGEX Venus YFP
+*PUC19
-<br>Membren Venus YFP
+*pGFPUV
-<br>PCR
+*pGEX Venus YFP
-<br>BBa_J63001①
+*Membren Venus YFP
+[[Team:Chiba/protocol/PCR|PCR]]
+*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]①
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
@@ Line 204: / Line 218: @@
 == 26,August,2008 ==
-<br>Gel check
+[[Team:Chiba/protocol/gelcheck|Gel Check]]
-<br>BBa_F2620
+*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
-<br>BBa_F2620
+*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
-<br>BBa_J63001
+*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
@@ Line 232: / Line 246: @@
 <br>Digestion
-<br>vector:BBa_F2620
+<br>vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
 <td width="257">Sample No</td>
-<td>BBa_F2620</td>
+<td>[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]</td>
 </tr>
 <tr>
@@ Line 304: / Line 318: @@
 <br>->Zymo Clean
 <br>Ligation
-<br>vector:BBa_F2620 insert:BBa_J52008①
+<br>vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:BBa_J52008①
-<br>vector:BBa_F2620 insert:BBa_E2030②
+<br>vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:BBa_E2030②
-<br>Negative control:BBa_F2620③
+<br>Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]③
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
@@ Line 337: / Line 351: @@
 </tr>
 </table>
-<br>->RT 2hour
-<br>->TRANSFORMATION XL10G
+-->RT 2hour
-<br>->37℃ over night
-<br>->colony PCR
+-->[[Team:Chiba/protocol/transformation|Transformation]](XL10G)
-<br>->MINI PREP
+-->37℃ over night
+-->colony PCR
+-->MINI PREP
 == 28,August,2008 ==
@@ Line 356: / Line 375: @@
 == 30,August,2008 ==
-<br>TRANSFORMATION
+<br>[[Team:Chiba/protocol/transformation|Transformation]]
 <br>mcherry
-<br>PCR
+<br>[[Team:Chiba/protocol/PCR|PCR]]
 <br>BBa_J52008(rLuc)
 <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">