Team:Chiba/Calendar-Home/26 August 2008

From 2008.igem.org

(Difference between revisions)
(→Team:Output)
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===Team:Output===
===Team:Output===
[[Team:Chiba/protocol/gelcheck|Gel Check]]
[[Team:Chiba/protocol/gelcheck|Gel Check]]
-
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
+
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1)
-
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
+
*[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2)
-
*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001]
+
*[http://partsregistry.org/Part:BBa_J63001 BBa_J63001](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>④</td><td>⑤</td>
+
<td>1</td><td>2</td><td>3</td>
</tr>
</tr>
<tr>
<tr>
<td>DNA tamplate</td>
<td>DNA tamplate</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>
</tr>
<tr>
<tr>
<td>Loading Dye</td>
<td>Loading Dye</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>
</tr>
<tr>
<tr>
<td>dH<sub>2</sub>O</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>
</tr>
</tr>
<tr>
<tr>
<td>TOTAL</td>
<td>TOTAL</td>
-
<td>6μl</td><td>6μl</td><td>6μl</td><td>6μl</td><td>6μl</td>
+
<td>6μl</td><td>6μl</td><td>6μl</td>
</tr>
</tr>
</table>
</table>

Latest revision as of 01:04, 30 October 2008

>Home | Notebook

25 August 2008 <|> 27 August 2008

Contents

Laboratory work

Team:Input

no work

Team:Communication

(8/25)--->Gel Check
Chiba-0825-2.JPG
Sample No. 1~3
Sample DNA(μL) 30
Loading Dye(μL) 6
TOTAL(μL) 36


1, [http://partsregistry.org/Part:BBa_C0178 BBa_C0178]
Insert:609bp -> OK
Chiba-0825-3.JPG
2, [http://partsregistry.org/Part:BBa_C0170 BBa_C0170]
Insert:609bp -> OK
Chiba-0825-4.JPG
3, [http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2007)
--> OK


--->Gel Extract


--->Zymo Clean
eluted with following amount of elution buffer
  1. 5μl
  2. 5μl
  3. 27μl


--->Gel Check
Chiba-0825-5.JPG
Sample No. 1~3
Sample DNA(μL) 2
Loading Dye(μL) 2
H2O(μL) 8
TOTAL(μL) 12
From left;
  1. insert:[http://partsregistry.org/Part:BBa_C0178 BBa_C0178] --> OK
  2. insert:[http://partsregistry.org/Part:BBa_C0170 BBa_C0170] --> OK?
  3. vector:[http://partsregistry.org/Part:BBa_J04500 BBa_J04500] --> OK


--->SAP
Sample No. 3
Sample DNA(μL) 26
SAP(μL) 1
SAP Buffer(μL) 3
TOTAL(μL) 30


--->Zymo Clean
Sample No.3 -->9μl


--->Gel Check
Chiba-0826.JPG
Sample No.3 1
Loading Dye(μL) 1
dH2O(μL) 4
TOTAL(μL) 6
Sample No.3 -> OK ･･･30ng?


--->Ligation
Sample No. (1)(2)(3)(4)(5)
insert(1)[http://partsregistry.org/Part:BBa_C0178 BBa_C0178](μL) 1--1-
insert(2)[http://partsregistry.org/Part:BBa_C0170 BBa_C0170](μL) -1--1
vector(3)[http://partsregistry.org/Part:BBa_J04500 BBa_J04500](μL) --222
ligase(μL) 11111
Buffer(μL) 22222
dH2O(μL) 66544
TOTAL(μL) 1010101010


--->Transformation
  1. insert(1)[http://partsregistry.org/Part:BBa_C0178 BBa_C0178]
  2. insert(2)[http://partsregistry.org/Part:BBa_C0170 BBa_C0170]
  3. vector(3)[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
  4. insert(1)[http://partsregistry.org/Part:BBa_C0178 BBa_C0178]+vector(3)[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]
  5. insert(2)[http://partsregistry.org/Part:BBa_C0170 BBa_C0170]+vector(3)[http://partsregistry.org/Part:BBa_J04500 BBa_J04500]

Team:Output

Gel Check

  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1)
  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620](2)
  • [http://partsregistry.org/Part:BBa_J63001 BBa_J63001](3)
Sample No. 123
DNA tamplate 111
Loading Dye 111
dH2O 444
TOTAL 6μl6μl6μl

Digestion

  • vector:[http://partsregistry.org/Part:BBa_F2620 BBa_F2620](1)
Sample No. 1
DNA tamplate 100
SpeI 2
PstI 2
BSA(x10) 13
Buffer2 13
TOTAL 130μl
  • insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](1)
[http://partsregistry.org/Part:BBa_E2030 BBa_E2030](2)
Sample No. 12
DNA tamplate 3030
XbaI 11
PstI 11
DpnI 11
Buffer3 44
BSA(x10) 44
TOTAL 40μl40μl


-->37°C　3hour
-->SAP
-->37°C 30min
-->65°C 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. 123
vector 555
inser 107.50
Ligase Buffer 444
Ligase 111
dH2O 02.510
TOTAL 20μl20μl20μl

-->RT 2hour

-->Transformation(XL10G)

-->37°C over night

-->colony PCR

-->Mini prep

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