Team:Chiba/jk/β/week 3

From 2008.igem.org

(Difference between revisions)

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

[http://partsregistry.org/Part:BBa_K084007 BBa_K084007(Plac+RBS+LasI)]
[http://partsregistry.org/Part:BBa_K084008 BBa_K084008(Plac+RBS+RhlI)]

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

Digestion

[http://partsregistry.org/Part:BBa_I9026 BBa_I9026](2007)
[http://partsregistry.org/Part:BBa_I9030 BBa_I9030](2006)
[http://partsregistry.org/Part:BBa_S03154 BBa_S03154](2007)
[http://partsregistry.org/Part:BBa_R0010 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
[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、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, [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①②	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 [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
Double Digestion Plac+RBS+RhlI 2~5
Double Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、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 [http://partsregistry.org/Part:BBa_S03154 BBa_S03154]-->

(2/9)Mini prepwith [http://partsregistry.org/Part:BBa_K084009 BBa_K084009], [http://partsregistry.org/Part:BBa_K084010 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.

[http://partsregistry.org/Part:BBa_K084009 BBa_K084009(Plac+RBS+RhlI+LVA, Amp)] -> 628 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
vector-4(Plac, Amp) -> 186 colonies

--->(2/9) Colony PCR

Transformation

Competent cells : JW1908 40μL

Transformed the following and grew on new ampicillin plates.

[http://partsregistry.org/Part:BBa_K084007 BBa_K084007(Plac+RBS+LasI)]
[http://partsregistry.org/Part:BBa_K084008 BBa_K084008(Plac+RBS+RhlI)]
[http://partsregistry.org/Part:BBa_K084010 BBa_T9002(Ptet+RBS+LuxR+GFP)]

--->(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)[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)).

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

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

[http://partsregistry.org/Part:BBa_K084007 BBa_K084007](Plac+RBS+LasI, Competent Cells : JW1908)
[http://partsregistry.org/Part:BBa_K084008 BBa_K084008](Plac+RBS+RhlI, Competent Cells : JW1908)
[http://partsregistry.org/Part:BBa_T9002 BBa_T9002](Ptet+RBS+LuxR+GFP, Competent Cells : JW1908)

from Glycerol Stock:

[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](Ptet+RBS+LuxI, Competent Cells : JW1908)

--->(3/9)Phenotype test

Transformation

Competent cells : XL10G 30μL

[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)

--->(4/9)Mini prep

3 September, 2008

(2/9)--->Mini prep

[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7)
[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8)
[http://partsregistry.org/Part:BBa_S03154 BBa_S03154]

--->Gel Check

Sample DNA	1
Loading Dye	1
dH₂O	4
TOTAL	6

From left;

[http://partsregistry.org/Part:BBa_S03154 BBa_S03154] -> OK 33ng/μL
[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7) -> OK

From left;

[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8) -> OK

--->Digestion test

[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7)
[http://partsregistry.org/Part:BBa_K084010 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 : [http://partsregistry.org/Part:BBa_K084009 BBa_K084009]R1,R3~R7 -> OK
Single Digestion : [http://partsregistry.org/Part:BBa_K084010 BBa_K084010]C1,C2,C4~C8 -> OK

From left;

Double Digestion : [http://partsregistry.org/Part:BBa_K084009 BBa_K084009]R1,R3~R7 -> OK
Double Digestion : [http://partsregistry.org/Part:BBa_K084010 BBa_K084010]C1,C2,C4~C8 -> OK

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

MIX

[http://partsregistry.org/Part:BBa_K084007 BBa_K084007](Plac+RBS+LasI, Competent Cells : JW1908) -> Sample Name : L1~L4
[http://partsregistry.org/Part:BBa_K084008 BBa_K084008](Plac+RBS+RhlI, Competent Cells : JW1908) -> Sample Name : R1~R4
[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](Ptet+RBS+LuxI, Competent Cells : JW1908)
[http://partsregistry.org/Part:BBa_T9002 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[http://partsregistry.org/Part:BBa_S03154]	-	-	-	-	-	-	-	-	2mL	-	-
AHL(100μM)	-	-	-	-	-	-	-	-	-	-	1μL
BBa_T9002[http://partsregistry.org/Part:BBa_T9002]	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;

[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007) -> OK

[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006) -> OK

From left;

[http://partsregistry.org/Part:BBa_C0161 BBa_C0261](2007) -> OK

[http://partsregistry.org/Part:BBa_C0161 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

[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)

ladder

Double digestion

[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)

Transformation

Competent cells : JW1908

Transformed the following and grew on new ampicillin plates.

[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R-3,4,6,7)
[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C-1,2,6,7)
[http://partsregistry.org/Part:BBa_T9002 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:

[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](Plac+RBS+RhlI+LVA, Competent Cells : JW1908)(r1, r3~r7)
[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](Plac+RBS+RhlI, Competent Cells : JW1908)(c1,c2,c4~c8)

from Glycerol Stock:

[http://partsregistry.org/Part:BBa_S03623 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[http://partsregistry.org/Part:BBa_S03154]	-	-	-	-	-	-	-	-	1mL	-	-
AHL(100μM)	-	-	-	-	-	-	-	-	-	-	1μL
BBa_T9002[http://partsregistry.org/Part:BBa_T9002]	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[http://partsregistry.org/Part:BBa_F2620](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 1: / Line 1: @@
 >[[Team:Chiba/jk/β|送受信班]]
+>[[Team:Chiba/protocol/j|Protocols]]
 ==Week 3==
@@ Line 7: / 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 14: / 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 50: / 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 67: / 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 92: / 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 109: / 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 121: / 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 131: / 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
 |}
 ===1 September, 2008===
@@ Line 144: / 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 202: / Line 208: @@
 <td>1</td></tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>4</td></tr>
 <tr>
@@ Line 212: / Line 218: @@
 ::*insert-1(I9026) -> OK
 ::*insert-2(I9030) -> OK
-::*insert-3(S03154) -> None
+::*insert-3(S03154) -> None --> Transformation [http://partsregistry.org/Part:BBa_S03154 BBa_S03154]-->
-::*vector-4(R0010) -> OK
+(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 220: / 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 244: / 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 256: / Line 263: @@
 :--->'''[[Team:Chiba/protocol/transformation|Transformation]]'''
 :Competent cells : XL10GOLD  30μL
-::*BBa_K084009(Plac+RBS+RhlI+LVA)[http://partsregistry.org/Part:BBa_K084009] : Sample No.L1~L4
+:Transformed the following and grew on new ampicillin plates.
-::*BBa_K084010(Plac+RBS+CinI+LVA)[http://partsregistry.org/Part:BBa_K084010] : Sample No.R1~R4
+::#[http://partsregistry.org/Part:BBa_K084009 BBa_K084009(Plac+RBS+RhlI+LVA, Amp)] -> 628 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
+::#vector-4(Plac, Amp) -> 186 colonies
+:--->(2/9)'''[[Team:Chiba/protocol/PCR| Colony PCR]]'''
+'''[[Team:Chiba/protocol/transformation|Transformation]]'''
+:Competent cells : JW1908  40μL
+:Transformed the following and grew on new ampicillin plates.
+::*[http://partsregistry.org/Part:BBa_K084007 BBa_K084007(Plac+RBS+LasI)]
+::*[http://partsregistry.org/Part:BBa_K084008 BBa_K084008(Plac+RBS+RhlI)]
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_T9002(Ptet+RBS+LuxR+GFP)]
+:--->(2/9)'''Liquid Culture'''
-:--->(2/9)OD
 ===2 September, 2008===
@@ Line 269: / Line 295: @@
 :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample DNA</td>
-<td>①</td><td>②</td><td>③</td><td>④</td>
+<td>1</td></tr>
-</tr>
-<tr>
-<td>Sample DNA</td>
-<td>3</td><td>3</td><td>3</td><td>3</td>
-</tr>
 <tr>
 <td>Loading Dye</td>
-<td>2</td><td>2</td><td>2</td><td>2</td>
+<td>1</td></tr>
-</tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
-<td>7</td><td>7</td><td>7</td><td>7</td>
+<td>4</td></tr>
-</tr>
 <tr>
 <td>TOTAL</td>
-<td>12</td><td>12</td><td>12</td><td>12</td>
+<td>6</td>
 </tr>
 </table>
 :From left;
-::I9026
+::I9026 -> OK, 100/μL
-::I9030
+::I9030 -> OK, 50ng/μL
-::S03154
+::S03154 -> OK, 30ng/μL (too low for the ligation:1/9 )
 |}
-:(1/9)--->'''[[Team:Chiba/protocol/PCR|Colony-PCR]]'''
+:(1/9)--->'''[[Team:Chiba/protocol/PCR| Colony PCR]]'''
-::Colony PCR of 8 colonies from ligation plates (1/9-(1),(2)) 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 326: / Line 345: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>20.5</td>
 </tr>
@@ Line 340: / Line 359: @@
 --->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
+:{|align="justify"
-{|align="justify"
 |[[Image:Chiba-0902-2.JPG]]
 |
@@ Line 353: / Line 371: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>4</td>
 </tr>
@@ Line 385: / 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 394: / Line 412: @@
-:(1/9)--->OD測
+(1/9)--->'''Liquid Culture'''
+:Cultured the following cells (2mL LB-Amp, at 37℃, 7 hours)
+::from transformed plates:
+::*[http://partsregistry.org/Part:BBa_K084007 BBa_K084007](Plac+RBS+LasI, Competent Cells : JW1908)
+::*[http://partsregistry.org/Part:BBa_K084008 BBa_K084008](Plac+RBS+RhlI, Competent Cells : JW1908)
+::*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002](Ptet+RBS+LuxR+GFP, Competent Cells : JW1908)
+::from Glycerol Stock:
+::*[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](Ptet+RBS+LuxI, Competent Cells : JW1908)
+--->(3/9)'''Phenotype test'''
@@ Line 400: / 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]]'''
+--->(4/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
 ===3 September, 2008===
 :(2/9)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7)
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8)
+::*[http://partsregistry.org/Part:BBa_S03154 BBa_S03154]
 :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
@@ Line 424: / Line 455: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>4</td>
 </tr>
@@ Line 433: / Line 464: @@
 </table>
 :From left;
-::*S03154 -> OK
+::*[http://partsregistry.org/Part:BBa_S03154 BBa_S03154] -> OK  33ng/μL
-::*R1,R3~R7  (Plac+RBS+RhlI+LVA) -> OK
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7) -> OK
 |}
@@ Line 441: / Line 472: @@
 |
 :From left;
-::*C1,C2,C4~C8  (Plac+RBS+CinI+LVA) -> OK
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8) -> OK
 |}
 :--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
-::*R1,R3~R7  (Plac+RBS+RhlI+LVA)
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R1, R3~R7)
-::*C1,C2,C4~C8  (Plac+RBS+CinI+LVA)
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C1,C2,C4~C8)
 <table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
@@ Line 478: / Line 509: @@
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
 <td>7</td><td>5</td></tr>
 <tr>
 <td>TOTAL</td>
-<td>10</td><td>10</td>
+<td>10μL</td><td>10μL</td>
 </tr>
 </table>
@@ Line 505: / Line 536: @@
 </table>
 :From left;
-:*Single Digestion : R1,R3~R7 -> OK
+:*Single Digestion : [http://partsregistry.org/Part:BBa_K084009 BBa_K084009]R1,R3~R7 -> OK
-:*Single Digestion : C1,C2,C4~C8 -> OK
+:*Single Digestion : [http://partsregistry.org/Part:BBa_K084010 BBa_K084010]C1,C2,C4~C8 -> OK
 |}
@@ Line 513: / Line 544: @@
 |
 :From left;
-:*Double Digestion : R1,R3~R7 -> OK
+:*Double Digestion : [http://partsregistry.org/Part:BBa_K084009 BBa_K084009]R1,R3~R7 -> OK
-:*Double Digestion : C1,C2,C4~C8 -> OK
+:*Double Digestion : [http://partsregistry.org/Part:BBa_K084010 BBa_K084010]C1,C2,C4~C8 -> OK
 |}
-(2/9)--->'''Phenotype-test : mix'''
+:(2/9)--->'''Phenotype-test'''
+:*MIX
+::*[http://partsregistry.org/Part:BBa_K084007 BBa_K084007](Plac+RBS+LasI, Competent Cells : JW1908) -> Sample Name : L1~L4
+::*[http://partsregistry.org/Part:BBa_K084008 BBa_K084008](Plac+RBS+RhlI, Competent Cells : JW1908) -> Sample Name : R1~R4
+::*[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](Ptet+RBS+LuxI, Competent Cells : JW1908)
+::*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002](Ptet+RBS+LuxR+GFP, Competent Cells : JW1908)
 :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
@@ Line 526: / Line 564: @@
 <tr>
 <td>L1</td>
-<td>2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td>
+<td>2mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td>
 </tr>
 <tr>
-<tr><td>L2</td><td>-</td><td>2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>L2</td><td>-</td><td>2mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
-<tr><td>L3</td><td>-</td><td>-</td><td>2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>L3</td><td>-</td><td>-</td><td>2mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
-<tr><td>L4</td><td>-</td><td>-</td><td>-</td><td>2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>L4</td><td>-</td><td>-</td><td>-</td><td>2mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
-<tr><td>R1</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>R1</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
-<tr><td>R2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>R2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
-<tr><td>R3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>R3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
-<tr><td>R4</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>R4</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>BBa_S03154[http://partsregistry.org/Part:BBa_S03154]</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>2mL</td><td>-</td><td>-</td></tr>
+<tr><td>AHL(100μM)</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1μL</td></tr>
+<tr><td>BBa_T9002[http://partsregistry.org/Part:BBa_T9002]</td><td>2mL</td><td>2mL</td><td>2mL</td><td>2mL</td><td>1mL</td><td>1mL</td><td>1mL</td><td>1mL</td><td>2mL</td><td>1mL</td><td>1mL</td></tr>
+<tr><td>IPTG(100μM)</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>-</td><td>-</td><td>-</td></tr>
+</table>
-<tr><td>BBa_S03154[http://partsregistry.org/Part:BBa_S03154]</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>2</td><td>-</td><td>-</td></tr>
+*Incubated for 8hours at 37 degrees
-<tr><td>BBa_T9002[http://partsregistry.org/Part:BBa_T9002]</td><td>2</td><td>2</td><td>2</td><td>2</td><td>1</td><td>1</td><td>1</td><td>1</td><td>2</td><td>1</td><td>1</td></tr>
+*Spindown (max rpm, 3 min)
-<tr><td>AHL</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td></tr>
+*The measurement of the intensity GFP(visual judgment)
+<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<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>
+<tr>
+<td>the intensity GFP</td>
+<td>+</td><td>+</td><td>+</td><td>+</td><td>+</td><td>+</td><td>+</td><td>++</td><td>+</td><td>+++</td><td>-</td>
+</tr>
 </table>
-:::|
-:::| 8h
-:::V
-::Spindown
-:::|
-:::|
-:::V
-::the intensity GFP
-===4 September, 2008===
+*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)
+<table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<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>
+<tr><td>the intensity GFP</td>
+<td>25.65</td><td>18.91</td><td>20.39</td><td>21.85</td><td>26.62</td><td>26.58</td><td>30.07</td><td>33.60</td><td>27.41</td><td>10.72</td><td>55.54</td>
+</tr>
+</table>
+===4 September, 2008===
 :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 {|align="justify"
@@ Line 558: / Line 615: @@
 :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample DNA</td>
-<td>①</td><td>②</td><td>③</td><td>④</td>
+<td>1</td>
-</tr>
-<tr>
-<td>Sample DNA</td>
-<td>3</td><td>3</td><td>3</td><td>3</td>
 </tr>
 <tr>
 <td>Loading Dye</td>
-<td>2</td><td>2</td><td>2</td><td>2</td>
+<td>1</td>
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
-<td>7</td><td>7</td><td>7</td><td>7</td>
+<td>4</td>
 </tr>
 <tr>
 <td>TOTAL</td>
-<td>12</td><td>12</td><td>12</td><td>12</td>
+<td>6μL</td>
 </tr>
 </table>
 :From left;
-:
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007) -> OK
-:
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007) -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006) -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006) -> OK
 |}
 {|align="justify"
 |[[Image:Chiba-0904-2.JPG]]
 |
-:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+:From left;
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0261](2007) -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0261](2007) -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0261](2006) -> OK
+::[http://partsregistry.org/Part:BBa_C0161 BBa_C0261](2006) -> OK
+:
+|}
+:--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
+<table width="600" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
-<td width="257">Sample No</td>
+<td width="257">Sample</td>
-<td>①</td><td>②</td><td>③</td><td>④</td>
+<td>Single Digesiton</td><td>Double Digestion</td>
 </tr>
 <tr>
-<td>Sample DNA</td>
+<td>Sample DNA</td><td>1</td><td>2</td>
-<td>3</td><td>3</td><td>3</td><td>3</td>
 </tr>
 <tr>
-<td>Loading Dye</td>
+<td>EcoRⅠ</td><td>0.05</td><td>0.05</td>
-<td>2</td><td>2</td><td>2</td><td>2</td>
+</tr>
+<tr>
+<td>PstⅠ</td><td>-</td><td>0.05</td>
+</tr>
+<tr>
+<td>Buffer 3</td>
+<td>1</td><td>1</td>
+</tr>
+<tr>
+<td>BSA</td>
+<td>1</td><td>1</td>
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
-<td>7</td><td>7</td><td>7</td><td>7</td>
+<td>6.95</td><td>5.9</td>
 </tr>
 <tr>
 <td>TOTAL</td>
-<td>12</td><td>12</td><td>12</td><td>12</td>
+<td>10μL</td><td>10μL</td>
 </tr>
 </table>
-:From left;
-:
-:
-|}
+:--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 {|align="justify"
 |[[Image:Chiba-0904-3.JPG]]
+:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample DNA</td>
+<td>10</td></tr>
+<tr>
+<td>Loading Dye</td>
+<td>2</td></tr>
+<tr>
+<td>TOTAL</td>
+<td>12μL</td></tr>
+</table>
 |
+:From left;
+::Single Digestion
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
+::ladder
+::Double digestion
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2007)
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
+::*[http://partsregistry.org/Part:BBa_C0161 BBa_C0161](2006)
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2007)
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
+::*[http://partsregistry.org/Part:BBa_C0261 BBa_C0261](2006)
+|}
+:'''[[Team:Chiba/protocol/transformation|Transformation]]'''
+:Competent cells : JW1908
+:Transformed the following and grew on new ampicillin plates.
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](R-3,4,6,7)
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](C-1,2,6,7)
+::*[http://partsregistry.org/Part:BBa_T9002 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:
+::*[http://partsregistry.org/Part:BBa_K084009 BBa_K084009](Plac+RBS+RhlI+LVA, Competent Cells : JW1908)(r1, r3~r7)
+::*[http://partsregistry.org/Part:BBa_K084010 BBa_K084010](Plac+RBS+RhlI, Competent Cells : JW1908)(c1,c2,c4~c8)
+::from Glycerol Stock:
+::*[http://partsregistry.org/Part:BBa_S03623 BBa_S03623](Ptet+RBS+LuxI, Competent Cells : JW1908)
+:--->'''Phenotype test'''
+:*MIX
 :<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>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>
 <tr>
-<td>Sample DNA</td>
+<td>r3</td>
-<td>3</td><td>3</td><td>3</td><td>3</td>
+<td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td>
+</tr>
+<tr>
+<tr><td>r4</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>r6</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>r7</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>c1</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>c2</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>c6</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>c7</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td><td>-</td></tr>
+<tr><td>BBa_S03154[http://partsregistry.org/Part:BBa_S03154]</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1mL</td><td>-</td><td>-</td></tr>
+<tr><td>AHL(100μM)</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1μL</td></tr>
+<tr><td>BBa_T9002[http://partsregistry.org/Part:BBa_T9002]</td><td>1mL</td><td>1mL</td><td>1mL</td><td>1mL</td><td>1mL</td><td>1mL</td><td>1mL</td><td>1mL</td><td>1mL</td><td>1mL</td><td>1mL</td></tr>
+<tr><td>IPTG(100μM)</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>1μL</td><td>-</td><td>-</td><td>-</td></tr>
+</table>
+*Incubated for 8hours at 37 degrees
+*The measurement of the intensity 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>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>
+<tr><td>the intensity GFP (Frist run)</td>
+<td>15.21</td><td>16.44</td><td>16.34</td><td>16.63</td><td>9.720</td><td>9.703</td><td>9.685</td><td>9.874</td><td>19.95</td><td>16.59</td><td>71.04</td>
+</tr>
+<tr><td>the intensity GFP (Second run)</td>
+<td>17.85</td><td>19.19</td><td>19.28</td><td>19.14</td><td>9.981</td><td>10.16</td><td>10.14</td><td>10.41</td><td>23.94</td><td>17.61</td><td>79.53</td>
+</tr>
+</table>
+*Spindown (max rpm, 3 min)
+*The measurement of the intensity GFP(visual judgment) (UV 312nm)
+<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<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>
+<tr>
+<td>the intensity GFP</td>
+<td>+</td><td>+</td><td>+</td><td>+</td><td>-</td><td>-</td><td>-</td><td>-</td><td>+</td><td>++</td><td>-</td>
+</tr>
+</table>
+===6 September, 2008===
+'''[[Team:Chiba/protocol/PCR|Colony-PCR]]'''
+::PCR of BBa_I9030 and two control,BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
+:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">DNA Template</td>
+<td>1</td>
+</tr>
+<tr>
+<td>dNTP mix</td>
+<td>5</td>
+</tr>
+<tr>
+<td>Foward Primer</td>
+<td>5</td>
+</tr>
+<tr>
+<td>Reverse Primer</td>
+<td>5</td>
+</tr>
+<tr>
+<td>DNA polymerase</td>
+<td>1</td>
+</tr>
+<tr>
+<td>Thermopol Buffer</td>
+<td>5</td>
+</tr>
+<tr>
+<td>dH<sub>2</sub>O</td>
+<td>33</td>
+</tr>
+<tr>
+<td>TOTAL</td>
+<td>50μL</td>
+</tr>
+</table>
+:95℃,5min -> ( 95℃,1min -> 54℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃ store
+:--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
+:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample DNA</td>
+<td>1</td>
 </tr>
 <tr>
 <td>Loading Dye</td>
-<td>2</td><td>2</td><td>2</td><td>2</td>
+<td>1</td>
 </tr>
 <tr>
-<td>dH2O</td>
+<td>dH<sub>2</sub>O</td>
-<td>7</td><td>7</td><td>7</td><td>7</td>
+<td>4</td>
 </tr>
 <tr>
 <td>TOTAL</td>
-<td>12</td><td>12</td><td>12</td><td>12</td>
+<td>6</td></tr>
+</table>
+:From left;<br>
+:1    I9030 without LVA  -> OK
+:2    I9030 Positive Control  -> OK
+:3    F2620 Positive control  -> OK
+'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
+Sample DNA : R0010
+:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<tr>
+<td width="257">Sample DNA</td>
+<td>1</td>
+</tr>
+<tr>
+<td>Loading Dye</td>
+<td>1</td>
+</tr>
+<tr>
+<td>dH<sub>2</sub>O</td>
+<td>4</td>
+</tr>
+<tr>
+<td>TOTAL</td>
+<td>6</td>
 </tr>
 </table>
-:From left;
-:
-:
-|}
-===5 September, 2008===
-===6 September, 2008===
+'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
+:Sample DNA:
+:R0010,2:S03154,3:J04500,4:C0161,5:C0261
+<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
+<td width="257">Sample No.</td>
+<td>1</td><td>2</td><td>3</td><td>4</td><td>5</td>
+<tr>
+<td>Sample DNA</td>
+<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
+</tr>
+<tr>
+<td>PstⅠ</td>
+<td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td><td>0.5</td>
+</tr>
+<tr>
+<td>SpeⅠ</td>
+<td>0.5</td><td>-</td><td>1</td><td>-</td><td>-</td>
+</tr>
+<tr>
+<td>XbaⅠ</td>
+<td>-</td><td>0.5</td><td>-</td><td>0.5</td><td>0.5</td>
+</tr>
+<tr>
+<td>Buffer 2</td>
+<td>2</td><td>-</td><td>3</td><td>-</td><td>-</td>
+</tr>
+<tr>
+<td>Buffer 3</td>
+<td>-</td><td>6</td><td>-</td><td>6</td><td>6</td>
+</tr>
+<tr>
+<td>(10×)BSA</td>
+<td>2</td><td>0.6</td><td>3</td><td>-</td><td>-</td>
+</tr>
+<tr>
+<td>(100×)BSA</td>
+<td>-</td><td>-</td><td>-</td><td>0.6</td><td>0.6</td>
+</tr>
+<tr>
+<td>dH<sub>2</sub>O</td>
+<td>3</td><td>6.4</td><td>7</td><td>1.4</td><td>1.4</td>
+</tr>
+<tr>
+<td>TOTAL</td>
+<td>20</td><td>60</td><td>30</td><td>60</td><td>60</td>
+</tr>
+</table>
 ===[[Team:Chiba/jk/β/week 4|>next week]]===
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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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