Team:Chiba/jk/β/week 5
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===19 September, 2008=== | ===19 September, 2008=== | ||
-->(?/9) | -->(?/9) | ||
+ | '''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
+ | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
+ | <tr> | ||
+ | <td width="257">Sample Name</td> | ||
+ | <td>J04500</td><td>R0010</td><td>C0161</td><td>Cin</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sample DNA</td> | ||
+ | <td>30</td><td>30</td><td>90</td><td>?</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Loading Dye</td> | ||
+ | <td>6</td><td>6</td><td>18</td><td>?</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>TOTAL</td> | ||
+ | <td>36</td><td>36</td><td>108</td><td>24</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | --->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''<br> | ||
+ | (各サンプル well3つずつ)<br> | ||
+ | *note:ADB buffer添加後,37℃,6h | ||
+ | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]''' | ||
+ | *elution | ||
+ | **J0450-->27 | ||
+ | **C0161-->11μL | ||
+ | --->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
+ | :--->'''[[Team:Chiba/protocol/ligation/ligation|Ligation]]''' | ||
+ | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
+ | <tr> | ||
+ | <td width="257">Sample Name</td> | ||
+ | <td>insert+vector</td><td>insert</td><td>vector</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>C0161</td> | ||
+ | <td>3</td><td>3</td><td>-</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>J04500</td> | ||
+ | <td>4.5</td><td>-</td><td>4.5</td> | ||
+ | </tr> | ||
+ | <td>ligase</td> | ||
+ | <td>1</td><td>1</td><td>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Buffer(invitrogen)</td> | ||
+ | <td>3</td><td>2</td><td>2</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>NFW?</td> | ||
+ | <td>3.5</td><td>4</td><td>2.5</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>TOTAL</td> | ||
+ | <td>15</td><td>10</td><td>10</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | :--->'''[[Team:Chiba/protocol/transformation|Transformation]]''' above ligation onto a LB-amp plate. | ||
+ | *note | ||
+ | **competent cell:XL10GOLD | ||
+ | **DNA:1μL,SOC:120μL | ||
- | + | --->(20/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]] | |
Line 68: | Line 129: | ||
===20 September, 2008=== | ===20 September, 2008=== | ||
+ | --->(19/9)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]] | ||
+ | |||
+ | --->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
+ | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
+ | <tr> | ||
+ | <td width="257">Sample Name</td> | ||
+ | <td>Lagation(J04500+C0161)</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sample DNA</td> | ||
+ | <td>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Loading Dye</td> | ||
+ | <td>1</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>H<sub>2</sub>O</td> | ||
+ | <td>4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>TOTAL</td> | ||
+ | <td>6</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | :From left; | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | |||
+ | :--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]''' | ||
+ | <table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
+ | <tr> | ||
+ | <td width="257">Sample No.</td> | ||
+ | <td>1-4</td><td>5-8</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sample DNA</td> | ||
+ | <td>3</td><td>3</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>XbaⅠ</td> | ||
+ | <td>0.25?</td><td>0.25</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>SpeⅠ</td> | ||
+ | <td>0.5?</td><td>0.5</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Buffer 2</td> | ||
+ | <td>2</td><td>2</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>BSA</td> | ||
+ | <td>2?</td><td>2?</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>dH<sub>2</sub>O</td> | ||
+ | <td>?</td><td>12.5</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>TOTAL</td> | ||
+ | <td>20</td><td>20</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | |||
+ | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
+ | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
+ | <tr> | ||
+ | <td width="257">Sample No.</td> | ||
+ | <td>1-4</td><td>5~8</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Sample DNA</td> | ||
+ | <td>20</td><td>20</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>Loading Dye</td> | ||
+ | <td>4</td><td>4</td> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td>TOTAL</td> | ||
+ | <td>24</td><td>24</td> | ||
+ | </tr> | ||
+ | </table> | ||
+ | :From left; | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | ::# -> OK | ||
+ | |||
+ | --->(?/9)'''[[Team:Chiba/protocol/transformation|Transformation]]''' | ||
+ | --->(22/9)'''phenotype check | ||
===[[Team:Chiba/jk/β/week 6|>next week]]=== | ===[[Team:Chiba/jk/β/week 6|>next week]]=== |
Latest revision as of 13:20, 1 October 2008
>送受信班
Contents |
Week 5
>last week
14 September, 2008
- (13/9)--->Mini prep
- Transformation
- Competent cells : XL10G
- BBa_T9002[http://partsregistry.org/Part:BBa_T9002](2007)
- Plac+RhlI+LVA
- Plac+CinI+LVA
- --->(15/9)
15 September, 2008
- Transformation
- competent cells : JW1908
- BBa_S03623 [http://partsregistry.org/Part:BBa_S03623]
- BBa_K084007 [http://partsregistry.org/Part:BBa_K084007]
- BBa_K084008 [http://partsregistry.org/Part:BBa_K084008]
- BBa_K084009 [http://partsregistry.org/Part:BBa_K084009]
- BBa_T9002 [http://partsregistry.org/Part:BBa_T9002]
(15/9)-->T9002 check(AHL sensitivity and time response test)
16 September, 2008
(15/9)->
- time-delay test
- conditions
- strain
Senders:JW1908
Receiver:JW1908
- temparature
37℃ and 30℃
17 September, 2008
- time-delay test
- conditions
- strain
Senders:JW1908
Receiver:JW1908
- temparature
37℃ and 30℃
- time-delay test
- conditions
- strain
Senders:XL10GOLD
Receiver:JW1908
- temparature
37℃ and 30℃
18 September, 2008
- time-delay test
- conditions
- strain
Senders:XL10GOLD
Receiver:JW1908
- temparature
37℃ and 30℃
19 September, 2008
-->(?/9) Gel Check
Sample Name | J04500 | R0010 | C0161 | Cin |
Sample DNA | 30 | 30 | 90 | ? |
Loading Dye | 6 | 6 | 18 | ? |
TOTAL | 36 | 36 | 108 | 24 |
--->Gel Extract
(各サンプル well3つずつ)
- note:ADB buffer添加後,37℃,6h
- --->Zymo Clean
- elution
- J0450-->27
- C0161-->11μL
--->Gel Check
- --->Ligation
Sample Name | insert+vector | insert | vector |
C0161 | 3 | 3 | - |
J04500 | 4.5 | - | 4.5 | ligase | 1 | 1 | 1 |
Buffer(invitrogen) | 3 | 2 | 2 |
NFW? | 3.5 | 4 | 2.5 |
TOTAL | 15 | 10 | 10 |
- --->Transformation 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℃ and Room Temparature
20 September, 2008
--->(19/9)Mini prep
--->Gel Check
Sample Name | Lagation(J04500+C0161) |
Sample DNA | 1 |
Loading Dye | 1 |
H2O | 4 |
TOTAL | 6 |
- From left;
- -> OK
- -> OK
- -> OK
- -> OK
- --->Digestion Test
Sample No. | 1-4 | 5-8 |
Sample DNA | 3 | 3 |
XbaⅠ | 0.25? | 0.25 |
SpeⅠ | 0.5? | 0.5 |
Buffer 2 | 2 | 2 |
BSA | 2? | 2? |
dH2O | ? | 12.5 |
TOTAL | 20 | 20 |
- --->Gel Check
Sample No. 1-4 5~8 Sample DNA 20 20 Loading Dye 4 4 TOTAL 24 24 - From left;
- -> OK
- -> OK
- -> OK
- -> OK
- -> OK
- -> OK
- -> OK
- -> OK
--->(?/9)Transformation
--->(22/9)phenotype check
>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 - From left;