Team:Chiba/Calendar-Home/19 September 2008

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Latest revision as of 00:20, 30 October 2008

>Home | Notebook

18 September 2008 <|> 20 September 2008

Laboratory work

Team:Communication

-->(?/9) Gel Check

Sample Name	[http://partsregistry.org/Part:BBa_J04500 J04500]	[http://partsregistry.org/Part:BBa_R0010 R0010]	[http://partsregistry.org/Part:BBa_C0161 C0161]	Cin
Sample DNA(μL)	30	30	90	20
Loading Dye(μL)	6	6	18	4
TOTAL(μL)	36	36	108	24

J04500' (Plac+RBS)

R0010 (Plac)

C0161 (RBS+Lux)

C0261 (Lux+LVA)

--->Gel Extract

(3 wells for 1 sample)

note:ADB buffer,
left at 37°C,for 6 hours

--->Zymo Clean

elution
- [http://partsregistry.org/Part:BBa_J04500 BBa_J04500]-->27μL
- [http://partsregistry.org/Part:BBa_C0161 BBa_C0161]-->11μL

--->Gel Check

--->Ligation

Sample Name	insert+vector	insert	vector
[http://partsregistry.org/Part:BBa_C0161 C0161](μL)	3	3	-
[http://partsregistry.org/Part:BBa_J04500 J04500](μL)	4.5	-	4.5
ligase(μL)	1	1	1
Buffer(invitrogen)(μL)	3	2	2
NFW(μL)	3.5	4	2.5
TOTAL(μL)	15	10	10

--->Transformed above ligation onto a LB-amp plate.

note
- competent cell:XL10GOLD
- DNA:1μL,SOC:120μL

--->(20/9)Mini prep

time-delay test
- conditions

strain
Senders:XL10GOLD
Receiver:JW1908
temparature

30°C and Room Temparature

Team:Output

colony PCR
insert:[http://partsregistry.org/Part:BBa_S03119 BBa_S03119](×16)(marker:Kan)
insert:[http://partsregistry.org/wiki/index.php?title=Part:BBa_K084004 K084004](×5)(marker:Kan)
insert:[http://partsregistry.org/wiki/index.php?title=Part:BBa_K084005 K084005](×7)(marker:Kan)
insert:[http://partsregistry.org/wiki/index.php?title=Part:BBa_K084006 K084006](×5)(marker:Kan)

@@ Line 7: / Line 7: @@
 -->(?/9)
 '''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
+{|align="justify"
+|[[Image:Chiba-0919.JPG|300px]]
+|
 <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
 <tr>
@@ Line 14: / Line 17: @@
 <tr>
 <td>Sample DNA(&mu;L)</td>
-<td>30</td><td>30</td><td>90</td><td>?</td>
+<td>30</td><td>30</td><td>90</td><td>20</td>
 </tr>
 <tr>
 <td>Loading Dye(&mu;L)</td>
-<td>6</td><td>6</td><td>18</td><td>?</td>
+<td>6</td><td>6</td><td>18</td><td>4</td>
 </tr>
 <tr>
@@ Line 25: / Line 28: @@
 </tr>
 </table>
+:
+::J04500' (Plac+RBS)
+|}
+[[Image:Chiba-0919-2.JPG|300px]] R0010 (Plac)
+[[Image:Chiba-0919-3.JPG|300px]] C0161 (RBS+Lux)
+[[Image:Chiba-0919-4.JPG|300px]] C0261 (Lux+LVA)
 --->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''<br>
 (3 wells for 1 sample)<br>
-*note:ADB buffer添加後,left at 37&deg;C,for 6 hours
+*note:ADB buffer,
+*left at 37&deg;C,for 6 hours
 :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
 *elution
@@ Line 82: / Line 98: @@
 &deg;C and Room Temparature
 ===Team:Output===