Team:Chiba/jk/β/week 3

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Latest revision as of 04:05, 24 October 2008

>送受信班

>Protocols

Week 3

>last week

31 August, 2008

Transformation

Competent Cells : XL10G

---> We saved these plates with a refrigerator.

Digestion

BBa_I9026(2007)
BBa_I9030(2006)
BBa_S03154(2007)
BBa_R0010(2007)

Sample No.	1~3	4
Sample DNA	12	10
PstⅠ	0.2	0.2
XbaⅠ	0.2	-
SpeⅠ	-	0.4
Buffer 2	-	1.5
Buffer 3	2	-
BSA	2	1.5
dH₂O	3.6	1.4
TOTAL	20	15

--->(1/9)Gel Check

(30/8)--->Mini prep

--->Digestion test

Plac+RBS+RhlI Sample No.2~5
BBa_T9002①、②

Sample	Single Digesiton 2~5	Double Digestion 2~5	Single Digestion T9002①	Single Digesiton T9002②	Double Digestion T9002①②
Sample DNA	1	3	3	1	3
XbaⅠ	0.1	0.1	0.1	0.1	0.1
SpeⅠ	-	0.1	-	-	0.1
Buffer 2	0.9	0.8	0.9	0.9	0.8
BSA	1	1	1	1	1
dH₂O	7	5	5	7	5
TOTAL	10	10	10	10	10

--->Gel Check

Sample No.	Plac+RBS+RhlI Sample No.2~5, BBa_T9002①②	Single & Double Digestion
Sample DNA	1	10
Loading Dye	1	2
dH₂O	4	-
TOTAL	6	12

From Left

Single Digestion Plac+RBS+RhlI 2~5
Single Digestion BBa_T9002①、②
Double Digestion Plac+RBS+RhlI 2~5
Double Digestion BBa_T9002①、②
BBa_T9002①、②

1 September, 2008

(31/8)--->Gel Check

Sample No.	1~3	4
Sample DNA	20	15
Loading Dye	4	3
TOTAL	24	18

From left;

insert-1(I9026)

insert-2(I9030)

insert-3(S03154)

vector-4(R0010)

--->Gel extract

--->zymo

insert-1(I9026) -> 7μL

insert-2(I9030) -> 7μL

insert-3(S03154) -> 7μL

vector-4(R0010) -> 15μL

--->SAP

vector-4(R0010)

--->Zymo

vector-4(R0010) -> 20μL

--->Gel Check

Sample DNA	1
Loading Dye	1
dH₂O	4
TOTAL	6

From left;

insert-1(I9026) -> OK
insert-2(I9030) -> OK
insert-3(S03154) -> None --> Transformation BBa_S03154-->

(2/9)Mini prepwith BBa_K084009, BBa_K084010

vector-4(R0010)

--->Ligation

Sample No.	(1)	(2)	(3)	(4)	(5)
insert-1(I9026)	3	-	3	-	-
insert-2(I9030)	-	3	-	3	-
vector-4(R0010)	3	3	-	-	3
ligase	1	1	1	1	1
Buffer	1	1	1	1	1
dH₂O	2	2	5	5	5
TOTAL	10	10	10	10	10

--->Transformation

Competent cells : XL10GOLD 30μL

Transformed the following and grew on new ampicillin plates.

BBa_K084009(Plac+RBS+RhlI+LVA, Amp) -> 628 colonies
BBa_K084010(Plac+RBS+CinI+LVA, Amp) -> 500 colonies
insert-1(RBS+RhlI+LVA) -> 9 colonies
insert-2(RBS+CinI+LVA) -> No colonies on the plate
vector-4(Plac, Amp) -> 186 colonies

--->(2/9) Colony PCR

Transformation

Competent cells : JW1908 40μL

Transformed the following and grew on new ampicillin plates.

--->(2/9)Liquid Culture

2 September, 2008

(31/8)--->Gel Check

Sample DNA	1
Loading Dye	1
dH₂O	4
TOTAL	6

From left;

I9026 -> OK, 100/μL

I9030 -> OK, 50ng/μL

S03154 -> OK, 30ng/μL (too low for the ligation:1/9 )

(1/9)---> Colony PCR

Colony PCR of 8 colonies from ligation plates (1/9:(1)BBa_K084009(R1~R8),(2)BBa_K084010(C1~C8)) and one from control plate(BBa_F2620(2007)).

DNA Template	1
dNTP mix	5
Foward Primer	0.3
Reverse Primer	0.3
DNA polymerase TAQ	0.5
Thermopol Buffer	3
dH₂O	20.5
TOTAL	30μL

95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃

--->Gel Check

Sample DNA	1
Loading Dye	1
dH₂O	4
TOTAL	6

From left;

Plac+RBS+RhlI+LVA

R1 -> OK

R2 -> Bad

R3~R7 -> OK

R8 -> Bad

From left;

Plac+RBS+CinI+LVA

C1,C2 -> OK

C3 -> Bad

C4~C6 -> OK

From left;

Plac+RBS+CinI+LVA

C7,C8 -> OK

BBa_F2620(2007):Positive control -> OK

--->(3/9)Mini prep

(1/9)--->Liquid Culture

Cultured the following cells (2mL LB-Amp, at 37℃, 7 hours)

from transformed plates:

BBa_K084007(Plac+RBS+LasI, Competent Cells : JW1908)
BBa_K084008(Plac+RBS+RhlI, Competent Cells : JW1908)
BBa_T9002(Ptet+RBS+LuxR+GFP, Competent Cells : JW1908)

from Glycerol Stock:

BBa_S03623(Ptet+RBS+LuxI, Competent Cells : JW1908)

--->(3/9)Phenotype test

Transformation

Competent cells : XL10G 30μL

BBa_C0161(2007)
BBa_C0161(2006)
BBa_C0261(2007)
BBa_C0261(2006)

--->(4/9)Mini prep

3 September, 2008

(2/9)--->Mini prep

BBa_K084009(R1, R3~R7)
BBa_K084010(C1,C2,C4~C8)
BBa_S03154

--->Gel Check

Sample DNA	1
Loading Dye	1
dH₂O	4
TOTAL	6

From left;

BBa_S03154 -> OK 33ng/μL
BBa_K084009(R1, R3~R7) -> OK

From left;

BBa_K084010(C1,C2,C4~C8) -> OK

--->Digestion test

BBa_K084009(R1, R3~R7)
BBa_K084010(C1,C2,C4~C8)

Digestion	Single	Double
Sample DNA	1	3
XbaⅠ	0.1	0.1
PstⅠ	0.1	0.1
Buffer 2	0.9	-
Buffer 3	-	-0.8
BSA	1	1
dH₂O	7	5
TOTAL	10μL	10μL

--->Gel Check

Sample DNA	10
Loading Dye	2
TOTAL	12

From left;

Single Digestion : BBa_K084009R1,R3~R7 -> OK
Single Digestion : BBa_K084010C1,C2,C4~C8 -> OK

From left;

Double Digestion : BBa_K084009R1,R3~R7 -> OK
Double Digestion : BBa_K084010C1,C2,C4~C8 -> OK

(2/9)--->Phenotype-test

MIX

BBa_K084007(Plac+RBS+LasI, Competent Cells : JW1908) -> Sample Name : L1~L4
BBa_K084008(Plac+RBS+RhlI, Competent Cells : JW1908) -> Sample Name : R1~R4
BBa_S03623(Ptet+RBS+LuxI, Competent Cells : JW1908)
BBa_T9002(Ptet+RBS+LuxR+GFP, Competent Cells : JW1908)

Sample No.	1	2	3	4	5	6	7	8	9	10	11
L1	2mL	-	-	-	-	-	-	-	-	-	-
L2	-	2mL	-	-	-	-	-	-	-	-	-
L3	-	-	2mL	-	-	-	-	-	-	-	-
L4	-	-	-	2mL	-	-	-	-	-	-	-
R1	-	-	-	-	1mL	-	-	-	-	-	-
R2	-	-	-	-	-	1mL	-	-	-	-	-
R3	-	-	-	-	-	-	1mL	-	-	-	-
R4	-	-	-	-	-	-	-	1mL	-	-	-
BBa_S03154[1]	-	-	-	-	-	-	-	-	2mL	-	-
AHL(100μM)	-	-	-	-	-	-	-	-	-	-	1μL
BBa_T9002[2]	2mL	2mL	2mL	2mL	1mL	1mL	1mL	1mL	2mL	1mL	1mL
IPTG(100μM)	1μL	1μL	1μL	1μL	1μL	1μL	1μL	1μL	-	-	-

Incubated for 8hours at 37 degrees
Spindown (max rpm, 3 min)
The measurement of the intensity GFP(visual judgment)

Sample No.	1	2	3	4	5	6	7	8	9	10	11
the intensity GFP	+	+	+	+	+	+	+	++	+	+++	-

Leaving for 12 at room temperture.
Resuspension
The measurement of the intensity of GFP by Fluoroskan Ascent 2.5(program:Ascent Software Version 2.6)

Sample No.	1	2	3	4	5	6	7	8	9	10	11
the intensity GFP	25.65	18.91	20.39	21.85	26.62	26.58	30.07	33.60	27.41	10.72	55.54

4 September, 2008

--->Gel Check

Sample DNA	1
Loading Dye	1
dH₂O	4
TOTAL	6μL

From left;

BBa_C0161(2007) -> OK

BBa_C0161(2006) -> OK

From left;

BBa_C0261(2007) -> OK

BBa_C0261(2006) -> OK

--->Digestion test

Sample	Single Digesiton	Double Digestion
Sample DNA	1	2
EcoRⅠ	0.05	0.05
PstⅠ	-	0.05
Buffer 3	1	1
BSA	1	1
dH₂O	6.95	5.9
TOTAL	10μL	10μL

--->Gel Check

Sample DNA	10
Loading Dye	2
TOTAL	12μL

From left;

Single Digestion

BBa_C0161(2007)
BBa_C0161(2007)
BBa_C0161(2006)
BBa_C0161(2006)
BBa_C0261(2007)
BBa_C0261(2007)
BBa_C0261(2006)
BBa_C0261(2006)

ladder

Double digestion

BBa_C0161(2007)
BBa_C0161(2007)
BBa_C0161(2006)
BBa_C0161(2006)
BBa_C0261(2007)
BBa_C0261(2007)
BBa_C0261(2006)
BBa_C0261(2006)

Transformation

Competent cells : JW1908

Transformed the following and grew on new ampicillin plates.

BBa_K084009(R-3,4,6,7)
BBa_K084010(C-1,2,6,7)
BBa_T9002(2007)

--->(5/9)Liquid Culture

5 September, 2008

(4/9)--->Liquid Culture

Cultured the following cells (2mL LB-Amp, at 37℃)

from transformed plates:

BBa_K084009(Plac+RBS+RhlI+LVA, Competent Cells : JW1908)(r1, r3~r7)
BBa_K084010(Plac+RBS+RhlI, Competent Cells : JW1908)(c1,c2,c4~c8)

from Glycerol Stock:

BBa_S03623(Ptet+RBS+LuxI, Competent Cells : JW1908)

--->Phenotype test

MIX

Sample No.	1	2	3	4	5	6	7	8	9	10	11
r3	1mL	-	-	-	-	-	-	-	-	-	-
r4	-	1mL	-	-	-	-	-	-	-	-	-
r6	-	-	1mL	-	-	-	-	-	-	-	-
r7	-	-	-	1mL	-	-	-	-	-	-	-
c1	-	-	-	-	1mL	-	-	-	-	-	-
c2	-	-	-	-	-	1mL	-	-	-	-	-
c6	-	-	-	-	-	-	1mL	-	-	-	-
c7	-	-	-	-	-	-	-	1mL	-	-	-
BBa_S03154[3]	-	-	-	-	-	-	-	-	1mL	-	-
AHL(100μM)	-	-	-	-	-	-	-	-	-	-	1μL
BBa_T9002[4]	1mL	1mL	1mL	1mL	1mL	1mL	1mL	1mL	1mL	1mL	1mL
IPTG(100μM)	1μL	1μL	1μL	1μL	1μL	1μL	1μL	1μL	-	-	-

Incubated for 8hours at 37 degrees
The measurement of the intensity GFP by Fluoroskan Ascent 2.5(program:Ascent Software Version 2.6)

Sample No.	1	2	3	4	5	6	7	8	9	10	11
the intensity GFP (Frist run)	15.21	16.44	16.34	16.63	9.720	9.703	9.685	9.874	19.95	16.59	71.04
the intensity GFP (Second run)	17.85	19.19	19.28	19.14	9.981	10.16	10.14	10.41	23.94	17.61	79.53

Spindown (max rpm, 3 min)
The measurement of the intensity GFP(visual judgment) (UV 312nm)

Sample No.	1	2	3	4	5	6	7	8	9	10	11
the intensity GFP	+	+	+	+	-	-	-	-	+	++	-

6 September, 2008

Colony-PCR

PCR of BBa_I9030 and two control,BBa_F2620[5](2007)).

DNA Template	1
dNTP mix	5
Foward Primer	5
Reverse Primer	5
DNA polymerase	1
Thermopol Buffer	5
dH₂O	33
TOTAL	50μL

95℃,5min -> ( 95℃,1min -> 54℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃ store

--->Gel Check

Sample DNA	1
Loading Dye	1
dH₂O	4
TOTAL	6

From left;

1 I9030 without LVA -> OK

2 I9030 Positive Control -> OK

3 F2620 Positive control -> OK

Gel Check Sample DNA : R0010

Sample DNA	1
Loading Dye	1
dH₂O	4
TOTAL	6

Digestion Test

Sample DNA:

1:R0010,2:S03154,3:J04500,4:C0161,5:C0261

Sample No.	1	2	3	4	5
Sample DNA	1	1	1	1	1
PstⅠ	0.5	0.5	0.5	0.5	0.5
SpeⅠ	0.5	-	1	-	-
XbaⅠ	-	0.5	-	0.5	0.5
Buffer 2	2	-	3	-	-
Buffer 3	-	6	-	6	6
(10×)BSA	2	0.6	3	-	-
(100×)BSA	-	-	-	0.6	0.6
dH₂O	3	6.4	7	1.4	1.4
TOTAL	20	60	30	60	60

>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 10: / Line 10: @@
 :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
 :Competent Cells : XL10G
-::*BBa_K084007(Plac+RBS+LasI)[http://partsregistry.org/Part:BBa_K084007]
+::*[http://partsregistry.org/Part:BBa_K084007 BBa_K084007(Plac+RBS+LasI)]
-::*BBa_K084008(Plac+RBS+RhlI)[http://partsregistry.org/Part:BBa_K084008]
+::*[http://partsregistry.org/Part:BBa_K084008 BBa_K084008(Plac+RBS+RhlI)]
 ::---> We saved these plates with a refrigerator.
@@ Line 17: / Line 17: @@
 :'''[[Team:Chiba/protocol/digestion|Digestion]]'''
-::#BBa_I9026[http://partsregistry.org/Part:BBa_I9026](2007)
+::#[http://partsregistry.org/Part:BBa_I9026 BBa_I9026](2007)
-::#BBa_I9030[http://partsregistry.org/Part:BBa_I9030](2006)
+::#[http://partsregistry.org/Part:BBa_I9030 BBa_I9030](2006)
-::#BBa_S03154[http://partsregistry.org/Part:BBa_S03154](2007)
+::#[http://partsregistry.org/Part:BBa_S03154 BBa_S03154](2007)
-::#BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
+::#[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
 :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample No.</td>
 <td>1~3</td><td>4</td>
 </tr>
@@ Line 53: / Line 53: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>3.6</td><td>1.4</td></tr>
 <tr>
@@ Line 70: / Line 70: @@
 :--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
 ::*Plac+RBS+RhlI Sample No.2~5
-::*BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
+::*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
 <table width="600" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
@@ Line 95: / Line 95: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>7</td><td>5</td><td>5</td><td>7</td><td>5</td>
 </tr>
@@ Line 112: / Line 112: @@
 :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample No.</td>
-<td>Plac+RBS+RhlI Sample No.2~5, BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①②
+<td>Plac+RBS+RhlI Sample No.2~5, [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①②
 </td><td>Single & Double Digestion</td>
 </tr>
@@ Line 124: / Line 124: @@
 <td>1</td><td>2</td></tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>4</td><td>-</td>
 </tr>
@@ Line 134: / Line 134: @@
 :From Left
 ::*Single Digestion Plac+RBS+RhlI 2~5
-::*Single Digestion BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
+::*Single Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
 ::*Double Digestion Plac+RBS+RhlI 2~5
-::*Double Digestion BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
+::*Double Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
-::*BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
+::*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
 |}
@@ Line 150: / Line 150: @@
 :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample No.</td>
 <td>1~3</td><td>4</td>
 </tr>
@@ Line 208: / Line 208: @@
 <td>1</td></tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>4</td></tr>
 <tr>
@@ Line 218: / Line 218: @@
 ::*insert-1(I9026) -> OK
 ::*insert-2(I9030) -> OK
-::*insert-3(S03154) -> None --> Transformation BBa_S03154[http://partsregistry.org/Part:BBa_S03154]-->
+::*insert-3(S03154) -> None --> Transformation [http://partsregistry.org/Part:BBa_S03154 BBa_S03154]-->
-(2/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''with BBa_K084009[http://partsregistry.org/Part:BBa_K084009], BBa_K084010[http://partsregistry.org/Part:BBa_K084010]
+(2/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''with [http://partsregistry.org/Part:BBa_K084009 BBa_K084009], [http://partsregistry.org/Part:BBa_K084010 BBa_K084010]
 ::*vector-4(R0010)
 |}
@@ Line 227: / Line 227: @@
 <table width="315" border="2" 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 251: / Line 251: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>2</td><td>2</td><td>5</td><td>5</td><td>5</td>
 </tr>
@@ Line 264: / Line 264: @@
 :Competent cells : XL10GOLD  30μL
 :Transformed the following and grew on new ampicillin plates.
-::#BBa_K084009(Plac+RBS+RhlI+LVA, Amp)[http://partsregistry.org/Part:BBa_K084009] -> 628 colonies
+::#[http://partsregistry.org/Part:BBa_K084009 BBa_K084009(Plac+RBS+RhlI+LVA, Amp)] -> 628 colonies
-::#BBa_K084010(Plac+RBS+CinI+LVA, Amp)[http://partsregistry.org/Part:BBa_K084010] -> 500 colonies
+::#[http://partsregistry.org/Part:BBa_K084010 BBa_K084010(Plac+RBS+CinI+LVA, Amp)] -> 500 colonies
 ::#insert-1(RBS+RhlI+LVA) -> 9 colonies
 ::#insert-2(RBS+CinI+LVA) -> No colonies on the plate
@@ Line 276: / Line 276: @@
 '''[[Team:Chiba/protocol/transformation|Transformation]]'''
-:Competent cells : BW ΔFliC  40μL
+:Competent cells : JW1908  40μL
 :Transformed the following and grew on new ampicillin plates.
-::*BBa_K084007(Plac+RBS+LasI)[http://partsregistry.org/Part:BBa_K084007]
+::*[http://partsregistry.org/Part:BBa_K084007 BBa_K084007(Plac+RBS+LasI)]
-::*BBa_K084008(Plac+RBS+RhlI)[http://partsregistry.org/Part:BBa_K084008]
+::*[http://partsregistry.org/Part:BBa_K084008 BBa_K084008(Plac+RBS+RhlI)]
-::*BBa_T9002(Ptet+RBS+LuxR+GFP)[http://partsregistry.org/Part:BBa_K084010]
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_T9002(Ptet+RBS+LuxR+GFP)]
 :--->(2/9)'''Liquid Culture'''
@@ Line 301: / Line 301: @@
 <td>1</td></tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>4</td></tr>
 <tr>
@@ Line 317: / Line 317: @@
 :(1/9)--->'''[[Team:Chiba/protocol/PCR| Colony PCR]]'''
-::Colony PCR of 8 colonies from ligation plates (1/9:(1)BBa_K084009[http://partsregistry.org/Part:BBa_K084009](R1~R8),(2)BBa_K084010[http://partsregistry.org/Part:BBa_K084010](C1~C8)) and one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
+::Colony PCR of 8 colonies from ligation plates (1/9:(1)[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1~R8),(2)[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1~C8)) and one from control plate([http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2007)).
 :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
@@ Line 345: / Line 345: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>20.5</td>
 </tr>
@@ Line 371: / Line 371: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>4</td>
 </tr>
@@ Line 403: / Line 403: @@
 :*Plac+RBS+CinI+LVA
 ::C7,C8 -> OK
-:*BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007):Positive control -> OK
+:*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2007):Positive control -> OK
 |}
@@ Line 415: / Line 415: @@
 :Cultured the following cells (2mL LB-Amp, at 37℃, 7 hours)
 ::from transformed plates:
-::*BBa_K084007(Plac+RBS+LasI, Competent Cells : BW ΔFliC)[http://partsregistry.org/Part:BBa_K084007]
+::*[http://partsregistry.org/Part:BBa_K084007 BBa_K084007](Plac+RBS+LasI, Competent Cells : JW1908)
-::*BBa_K084008(Plac+RBS+RhlI, Competent Cells : BW ΔFliC)[http://partsregistry.org/Part:BBa_K084008]
+::*[http://partsregistry.org/Part:BBa_K084008 BBa_K084008](Plac+RBS+RhlI, Competent Cells : JW1908)
-::*BBa_T9002(Ptet+RBS+LuxR+GFP, Competent Cells : BW ΔFliC)[http://partsregistry.org/Part:BBa_T9002]
+::*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002](Ptet+RBS+LuxR+GFP, Competent Cells : JW1908)
 ::from Glycerol Stock:
-::*BBa_S03623(Ptet+RBS+LuxI, Competent Cells : BW ΔFliC)[http://partsregistry.org/Part:BBa_S03623]
+::*[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](Ptet+RBS+LuxI, Competent Cells : JW1908)
 --->(3/9)'''Phenotype test'''
@@ Line 427: / Line 427: @@
 '''[[Team:Chiba/protocol/transformation|Transformation]]'''
 :Competent cells : XL10G  30μL
-::*C0161(2007) [http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
-::*C0161(2006) [http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
-::*C0261(2007) [http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
-::*C0261(2006) [http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
 --->(4/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
@@ Line 436: / Line 436: @@
 ===3 September, 2008===
 :(2/9)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
-::*BBa_K084009[http://partsregistry.org/Part:BBa_K084009](R1, R3~R7)
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7)
-::*BBa_K084010[http://partsregistry.org/Part:BBa_K084010](C1,C2,C4~C8)
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8)
-::*BBa_S03154[http://partsregistry.org/Part:BBa_S03154]
+::*[http://partsregistry.org/Part:BBa_S03154 BBa_S03154]
@@ Line 455: / Line 455: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>4</td>
 </tr>
@@ Line 464: / Line 464: @@
 </table>
 :From left;
-::*S03154[http://partsregistry.org/Part:BBa_S03154] -> OK  33ng/μL
+::*[http://partsregistry.org/Part:BBa_S03154 BBa_S03154] -> OK  33ng/μL
-::*BBa_K084009[http://partsregistry.org/Part:BBa_K084009](R1, R3~R7) -> OK
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7) -> OK
 |}
@@ Line 472: / Line 472: @@
 |
 :From left;
-::*BBa_K084010[http://partsregistry.org/Part:BBa_K084010](C1,C2,C4~C8) -> OK
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8) -> OK
 |}
 :--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
-::*BBa_K084009[http://partsregistry.org/Part:BBa_K084009](R1, R3~R7)
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7)
-::*BBa_K084010[http://partsregistry.org/Part:BBa_K084010](C1,C2,C4~C8)
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8)
 <table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
@@ Line 509: / Line 509: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>7</td><td>5</td></tr>
 <tr>
@@ Line 536: / Line 536: @@
 </table>
 :From left;
-:*Single Digestion : BBa_K084009[http://partsregistry.org/Part:BBa_K084009]R1,R3~R7 -> OK
+:*Single Digestion : [http://partsregistry.org/Part:BBa_K084009 BBa_K084009]R1,R3~R7 -> OK
-:*Single Digestion : BBa_K084010[http://partsregistry.org/Part:BBa_K084010]C1,C2,C4~C8 -> OK
+:*Single Digestion : [http://partsregistry.org/Part:BBa_K084010 BBa_K084010]C1,C2,C4~C8 -> OK
 |}
@@ Line 544: / Line 544: @@
 |
 :From left;
-:*Double Digestion : BBa_K084009[http://partsregistry.org/Part:BBa_K084009]R1,R3~R7 -> OK
+:*Double Digestion : [http://partsregistry.org/Part:BBa_K084009 BBa_K084009]R1,R3~R7 -> OK
-:*Double Digestion : BBa_K084010[http://partsregistry.org/Part:BBa_K084010]C1,C2,C4~C8 -> OK
+:*Double Digestion : [http://partsregistry.org/Part:BBa_K084010 BBa_K084010]C1,C2,C4~C8 -> OK
 |}
@@ Line 551: / Line 551: @@
 :(2/9)--->'''Phenotype-test'''
 :*MIX
-::*BBa_K084007(Plac+RBS+LasI, Competent Cells : BW ΔFliC)[http://partsregistry.org/Part:BBa_K084007] -> Sample Name : L1~L4
+::*[http://partsregistry.org/Part:BBa_K084007 BBa_K084007](Plac+RBS+LasI, Competent Cells : JW1908) -> Sample Name : L1~L4
-::*BBa_K084008(Plac+RBS+RhlI, Competent Cells : BW ΔFliC)[http://partsregistry.org/Part:BBa_K084008] -> Sample Name : R1~R4
+::*[http://partsregistry.org/Part:BBa_K084008 BBa_K084008](Plac+RBS+RhlI, Competent Cells : JW1908) -> Sample Name : R1~R4
-::*BBa_S03623(Ptet+RBS+LuxI, Competent Cells : BW ΔFliC)[http://partsregistry.org/Part:BBa_S03623]
+::*[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](Ptet+RBS+LuxI, Competent Cells : JW1908)
-::*BBa_T9002(Ptet+RBS+LuxR+GFP, Competent Cells : BW ΔFliC)[http://partsregistry.org/Part:BBa_T9002]
+::*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002](Ptet+RBS+LuxR+GFP, Competent Cells : JW1908)
@@ Line 585: / Line 585: @@
 <table width="315" border="2" 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><td>6</td><td>7</td><td>8</td><td>9</td><td>10</td><td>11</td>
 </tr>
@@ Line 597: / Line 597: @@
 *Leaving for 12 at room temperture.
 *Resuspension
-*The measurement of the intensity GFP by Fluoroskan Ascent 2.5(program:Ascent Software Version 2.6)
+*The measurement of the intensity of GFP by Fluoroskan Ascent 2.5(program:Ascent Software Version 2.6)
 <table width="500" border="2" 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><td>6</td><td>7</td><td>8</td><td>9</td><td>10</td><td>11</td>
 </tr>
@@ Line 623: / Line 623: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>4</td>
 </tr>
@@ Line 632: / Line 632: @@
 </table>
 :From left;
-::BBa_C0161(2007)[http://partsregistry.org/Part:BBa_C0161] -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007) -> OK
-::BBa_C0161(2007)[http://partsregistry.org/Part:BBa_C0161] -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007) -> OK
-::BBa_C0161(2006)[http://partsregistry.org/Part:BBa_C0161] -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006) -> OK
-::BBa_C0161(2006)[http://partsregistry.org/Part:BBa_C0161] -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006) -> OK
 |}
 {|align="justify"
@@ Line 641: / Line 641: @@
 |
 :From left;
-::BBa_C0261(2007)[http://partsregistry.org/Part:BBa_C0161] -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0261](2007) -> OK
-::BBa_C0261(2007)[http://partsregistry.org/Part:BBa_C0161] -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0261](2007) -> OK
-::BBa_C0261(2006)[http://partsregistry.org/Part:BBa_C0161] -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0261](2006) -> OK
-::BBa_C0261(2006)[http://partsregistry.org/Part:BBa_C0161] -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0261](2006) -> OK
 :
 |}
@@ Line 674: / Line 674: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>6.95</td><td>5.9</td>
 </tr>
@@ Line 703: / Line 703: @@
 :From left;
 ::Single Digestion
-::*BBa_C0161(2007)[http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
-::*BBa_C0161(2007)[http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
-::*BBa_C0161(2006)[http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
-::*BBa_C0161(2006)[http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
-::*BBa_C0261(2007)[http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
-::*BBa_C0261(2007)[http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
-::*BBa_C0261(2006)[http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
-::*BBa_C0261(2006)[http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
 ::ladder
 ::Double digestion
-::*BBa_C0161(2007)[http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
-::*BBa_C0161(2007)[http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
-::*BBa_C0161(2006)[http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
-::*BBa_C0161(2006)[http://partsregistry.org/Part:BBa_C0161]
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
-::*BBa_C0261(2007)[http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
-::*BBa_C0261(2007)[http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
-::*BBa_C0261(2006)[http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
-::*BBa_C0261(2006)[http://partsregistry.org/Part:BBa_C0261]
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
 |}
@@ Line 726: / Line 726: @@
 :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
-:Competent cells : BW ΔFliC
+:Competent cells : JW1908
 :Transformed the following and grew on new ampicillin plates.
-::*BBa_K084009[http://partsregistry.org/Part:BBa_K084009](R-3,4,6,7)
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R-3,4,6,7)
-::*BBa_K084010[http://partsregistry.org/Part:BBa_K084010](C-1,2,6,7)
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C-1,2,6,7)
-::*BBa_T9002(2007)[http://partsregistry.org/Part:BBa_T9002]
+::*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002](2007)
 :--->(5/9)'''Liquid Culture'''
@@ Line 738: / Line 738: @@
 :Cultured the following cells (2mL LB-Amp, at 37℃)
 ::from transformed plates:
-::*BBa_K084009(Plac+RBS+RhlI+LVA, Competent Cells : JW1908) [http://partsregistry.org/Part:BBa_K084009](r1, r3~r7)
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](Plac+RBS+RhlI+LVA, Competent Cells : JW1908)(r1, r3~r7)
-::*BBa_K084010(Plac+RBS+RhlI, Competent Cells : JW1908)[http://partsregistry.org/Part:BBa_K084010](c1,c2,c4~c8)
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](Plac+RBS+RhlI, Competent Cells : JW1908)(c1,c2,c4~c8)
 ::from Glycerol Stock:
-::*BBa_S03623(Ptet+RBS+LuxI, Competent Cells : JW1908)[http://partsregistry.org/Part:BBa_S03623]
+::*[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](Ptet+RBS+LuxI, Competent Cells : JW1908)
 :--->'''Phenotype test'''
@@ Line 773: / Line 773: @@
 <table width="500" border="2" 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><td>6</td><td>7</td><td>8</td><td>9</td><td>10</td><td>11</td>
 </tr>
@@ Line 790: / Line 790: @@
 <table width="315" border="2" 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><td>6</td><td>7</td><td>8</td><td>9</td><td>10</td><td>11</td>
 </tr>
@@ Line 829: / Line 829: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>33</td>
 </tr>
@@ Line 895: / Line 895: @@
 <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
-<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>

Team:Chiba/jk/β/week 3

From 2008.igem.org

Latest revision as of 04:05, 24 October 2008

Contents

Week 3

31 August, 2008

1 September, 2008

2 September, 2008

3 September, 2008

4 September, 2008

5 September, 2008

6 September, 2008

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