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From 2008.igem.org

Revision as of 05:38, 24 October 2008 (view source) Masahiro (Talk | contribs) ← Older edit		Revision as of 05:47, 24 October 2008 (view source) Masahiro (Talk | contribs) Newer edit →
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 ==
Line 96:		Line 97:
	*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②		*[http://partsregistry.org/Part:BBa_J33202 BBa_J33202]②
	*BL21③		*BL21③
-	*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]8④	+	*[http://partsregistry.org/Part:BBa_J52008 BBa_J52008]④
	*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤		*[http://partsregistry.org/Part:BBa_I712019 BBa_I712019]⑤
Line 179:		Line 180:
	<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 224:		Line 225:
	<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 245:		Line 246:
	</table>		</table>
-	<br>Digestion	+	[[Team:Chiba/protocol/digestion|Digestion]]
-	<br>vector:[http://partsregistry.org/Part:BBa_F2620 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>[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]</td>		<td>[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]</td>
	</tr>		</tr>
Line 277:		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 312:		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>-->Zymo Clean
-	<br>Ligation	+
-	<br>vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:BBa_J52008①	+	[[Team:Chiba/protocol/ligation/ligation|Ligation]]
-	<br>vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620] insert:BBa_E2030②	+
-	<br>Negative control:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]③	+	*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>
Line 360:		Line 367:
	-->colony PCR		-->colony PCR
-	-->MINI PREP	+	-->[[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 370:		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>[[Team:Chiba/protocol/transformation|Transformation]]	+	[[Team:Chiba/protocol/transformation|Transformation]]
-	<br>mcherry	+
-	<br>[[Team:Chiba/protocol/PCR|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 422:		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]]

---

Revision as of 05:47, 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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