"

 

From 2008.igem.org

Revision as of 12:16, 23 October 2008 (view source) Mai (Talk | contribs) (→30,August,2008) ← Older edit		Latest revision as of 05:50, 24 October 2008 (view source) Masahiro (Talk | contribs) (→26,August,2008)
(3 intermediate revisions not shown)
Line 1:		Line 1:
	[[Team:Chiba/jk|>実験ログ]]		[[Team:Chiba/jk|>実験ログ]]
-	<br>[[Team:Chiba/jk/γ|>出力班]]	+
		+	[[Team:Chiba/jk/γ|>出力班]]
	== WEEK 2 ==		== WEEK 2 ==
	== 24,August 2008 ==		== 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">		<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>①</td><td>②</td><td>③</td>		<td>①</td><td>②</td><td>③</td>
	</tr>		</tr>
Line 26:		Line 28:
	</tr>		</tr>
	<tr>		<tr>
-	<td>dH2O</td>	+	<td>dH<sub>2</sub>O</td>
	<td>6</td><td>6</td><td>6</td>		<td>6</td><td>6</td><td>6</td>
	</tr>		</tr>
Line 38:		Line 40:
	</tr>		</tr>
	</table>		</table>
-	<br>->37℃ 30min	+
		+
		+	-->37℃ 30min
		+
		+
	<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>①(afetr digestion)</td><td>②(after digestion)</td><td>③(afetr digestion)</td>		<td>①(afetr digestion)</td><td>②(after digestion)</td><td>③(afetr digestion)</td>
	</tr>		</tr>
Line 57:		Line 63:
	</tr>		</tr>
	</table>		</table>
		+
		+
	<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>①</td><td>②</td><td>③</td>		<td>①</td><td>②</td><td>③</td>
	</tr>		</tr>
Line 71:		Line 79:
	</tr>		</tr>
	<tr>		<tr>
-	<td>dH2O</td>	+	<td>dH<sub>2</sub>O</td>
	<td>3</td><td>3</td><td>3</td>		<td>3</td><td>3</td><td>3</td>
	</tr>		</tr>
Line 79:		Line 87:
	</tr>		</tr>
	</table>		</table>
-	<br>->135V 30min	+
		+	-->135V 30min
	== 25,August,2008 ==		== 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]④
-	<br>BBa_I712019⑤	+	*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤
		+
	<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>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>		<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
	</tr>		</tr>
Line 120:		Line 130:
	</tr>		</tr>
	<tr>		<tr>
-	<td>dH2O</td>	+	<td>dH<sub>2</sub>O</td>
	<td>68</td><td>68</td><td>68</td><td>68</td><td>68</td>		<td>68</td><td>68</td><td>68</td><td>68</td><td>68</td>
	</tr>		</tr>
Line 128:		Line 138:
	</tr>		</tr>
	</table>		</table>
-	<br>->Gel	+
		+	-->[[Team:Chiba/protocol/gelcheck|Gel Check]]
		+
		+
	<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>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>		<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
	</tr>		</tr>
Line 143:		Line 156:
	</tr>		</tr>
	<tr>		<tr>
-	<td>dH2O</td>	+	<td>dH<sub>2</sub>O</td>
	<td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>		<td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
	</tr>		</tr>
Line 152:		Line 165:
	</table>		</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">		<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>①</td>		<td>①</td>
	</tr>		</tr>
Line 193:		Line 208:
	</tr>		</tr>
	<tr>		<tr>
-	<td>dH2O</td>	+	<td>dH<sub>2</sub>O</td>
	<td>68</td>		<td>68</td>
	</tr>		</tr>
Line 204:		Line 219:
	== 26,August,2008 ==		== 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">		<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>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>		<td>①</td><td>②</td><td>③</td><td>④</td><td>⑤</td>
	</tr>		</tr>
Line 222:		Line 237:
	</tr>		</tr>
	<tr>		<tr>
-	<td>dH2O</td>	+	<td>dH<sub>2</sub>O</td>
	<td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>		<td>4</td><td>4</td><td>4</td><td>4</td><td>4</td>
	</tr>		</tr>
Line 231:		Line 246:
	</table>		</table>
-	<br>Digestion	+	[[Team:Chiba/protocol/digestion|Digestion]]
-	<br>vector:BBa_F2620	+	*vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
		+
	<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>BBa_F2620</td>	+	<td>[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]</td>
	</tr>		</tr>
	<tr>		<tr>
Line 263:		Line 279:
	</tr>		</tr>
	</table>		</table>
-	<br>insert:BBa_J52008,BBa_E2030	+
		+	*insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008],[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]
		+
	<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>J52008</td><td>E2030</td>	+	<td>[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]</td><td>[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]</td>
	</tr>		</tr>
	<tr>		<tr>
Line 298:		Line 316:
	</tr>		</tr>
	</table>		</table>
-	<br>->37℃　3hour	+	<br>-->37℃　3hour
-	<br>->SAP	+	<br>-->SAP
-	<br>->37℃ 30min	+	<br>-->37℃ 30min
-	<br>->65℃ 15min	+	<br>-->65℃ 15min
-	<br>->Zymo Clean	+	<br>-->[[Team:Chiba/protocol/DNA_Purification/zymocleam|Zymo Clean]]
-	<br>Ligation	+
-	<br>vector:BBa_F2620 insert:BBa_J52008①	+	[[Team:Chiba/protocol/ligation/ligation|Ligation]]
-	<br>vector:BBa_F2620 insert:BBa_E2030②	+
-	<br>Negative control:BBa_F2620③	+	*vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
		+	*vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]②
		+	*Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]③
		+
	<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>①</td><td>②</td><td>③</td>		<td>①</td><td>②</td><td>③</td>
	</tr>		</tr>
Line 329:		Line 350:
	</tr>		</tr>
	<tr>		<tr>
-	<td>dH2O</td>	+	<td>dH<sub>2</sub>O</td>
	<td>0</td><td>2.5</td><td>10</td>		<td>0</td><td>2.5</td><td>10</td>
	</tr>		</tr>
Line 337:		Line 358:
	</tr>		</tr>
	</table>		</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
		+
		+	-->[[Team:Chiba/protocol/DNA_Purification/sigma|Mini prep]]
	== 28,August,2008 ==		== 28,August,2008 ==
-	<br>[[Team:Chiba/jk/γ/Trl|Time Responce(liquid)]]	+
		+	[[Team:Chiba/jk/γ/Trl|Time Responce(liquid)]]
Line 351:		Line 378:
	== 29,August,2008 ==		== 29,August,2008 ==
-	<br>[[Team:Chiba/jk/γ/trs|TIME RESPONCE(solid)]]	+
		+	[[Team:Chiba/jk/γ/trs|TIME RESPONCE(solid)]]
	== 30,August,2008 ==		== 30,August,2008 ==
-	<br>TRANSFORMATION	+	[[Team:Chiba/protocol/transformation|Transformation]]
-	<br>mcherry	+
-	<br>PCR	+	*mcherry
-	<br>BBa_J52008(rLuc)	+
		+	[[Team:Chiba/protocol/PCR|PCR]]
		+
		+	*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](rLuc)
		+
	<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>BBa_J52008</td>	+	<td>[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]</td>
	</tr>		</tr>
	<tr>		<tr>
Line 390:		Line 422:
	</tr>		</tr>
	<tr>		<tr>
-	<td>dH2O</td>	+	<td>dH<sub>2</sub>O</td>
	<td>34</td>		<td>34</td>
	</tr>		</tr>
Line 403:		Line 435:
	[[Team:Chiba/jk/γ/week 1|>Last week]]		[[Team:Chiba/jk/γ/week 1|>Last week]]
-	<br>[[Team:Chiba/jk/γ/week 3|>Next week]]	+
		+	[[Team:Chiba/jk/γ/week 3|>Next week]]

---

Latest revision as of 05:50, 24 October 2008

>実験ログ

>出力班

Contents 1 WEEK 2 2 24,August 2008 3 25,August,2008 4 26,August,2008 5 28,August,2008 6 29,August,2008 7 30,August,2008

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]④
• [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:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008],[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]

Sample No.	[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]	[http://partsregistry.org/Part:BBa_E2030 BBa_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:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]①
• vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:[http://partsregistry.org/Part:BBa_E2030 BBa_E2030]②
• 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

• [http://partsregistry.org/Part:BBa_J52008 BBa_J52008](rLuc)

Sample No.	[http://partsregistry.org/Part:BBa_J52008 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

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