"
Team:Chiba/Calendar-Home/25 September 2008
From 2008.igem.org
(→Team:Output) |
|||
| Line 8: | Line 8: | ||
===Team:Output=== | ===Team:Output=== | ||
<br>colony PCR | <br>colony PCR | ||
| - | <br>Vector:BBa_R0062 Insert:BBa_E0240 | + | <br>Vector:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] Insert:[http://partsregistry.org/Part:BBa_E0240 BBa_E0240] |
| - | <br>Vector:BBa_R0062 | + | <br>Vector:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
| Line 52: | Line 52: | ||
<br>Digestion Test | <br>Digestion Test | ||
| - | <br>BBa_I6206 | + | <br>[http://partsregistry.org/Part:BBa_I6206 BBa_I6206] |
| - | <br>BBa_I6101 | + | <br>[http://partsregistry.org/Part:BBa_I6101 BBa_I6101] |
| - | <br>BBa_I6042 | + | <br>[http://partsregistry.org/Part:BBa_I6042 BBa_I6042] |
| - | <br>BBa_I0462 | + | <br>[http://partsregistry.org/Part:BBa_I0462 BBa_I0462] |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
| Line 85: | Line 85: | ||
<br>->30 min 37°C | <br>->30 min 37°C | ||
<br>Gel check | <br>Gel check | ||
| - | <br>BBa_I6206(digestion product) | + | <br>[http://partsregistry.org/Part:BBa_I6206 BBa_I6206](digestion product) |
| - | <br>BBa_I6101(digestion product) | + | <br>[http://partsregistry.org/Part:BBa_I6101 BBa_I6101](digestion product) |
| - | <br>BBa_I6042(digestion product) | + | <br>[http://partsregistry.org/Part:BBa_I6042 BBa_I6042](digestion product) |
| - | <br>BBa_I0462(digestion product) | + | <br>[http://partsregistry.org/Part:BBa_I0462 BBa_I0462](digestion product) |
| - | <br>BBa_I0642(natural) | + | <br>[http://partsregistry.org/Part:BBa_I0642 BBa_I0642](natural) |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
| Line 113: | Line 113: | ||
<br>Digestion | <br>Digestion | ||
| - | <br>BBa_I0462 | + | <br>[http://partsregistry.org/Part:BBa_I0462 BBa_I0462] |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
| Line 149: | Line 149: | ||
<br>Ligation | <br>Ligation | ||
| - | <br>Vector:BBa_I7100 | + | <br>Vector:[http://partsregistry.org/Part:BBa_I7100 BBa_I7100] |
| - | <br>Insert:BBa_R0062 | + | <br>Insert:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] |
| - | <br>Insert:BBa_I0462 | + | <br>Insert:[http://partsregistry.org/Part:BBa_I0462 BBa_I0462] |
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | <table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
| Line 174: | Line 174: | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| - | <td>dH<sub>2</sub>O(μL)<td> | + | <td>dH<sub>2</sub>O(μL)</td> |
<td>0.4</td><td>5</td> | <td>0.4</td><td>5</td> | ||
</tr> | </tr> | ||
Revision as of 23:20, 29 October 2008
24 September 2008 <|> 26 September 2008
Laboratory work
Team:Communication
Team:Output
colony PCR
Vector:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062] Insert:[http://partsregistry.org/Part:BBa_E0240 BBa_E0240]
Vector:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]
| Sample No. | each |
| culture(μL) | 1 |
| Fwd primer(μL) | 1.5 |
| Rev primer(μL) | 1.5 |
| Taq(DNA pol)(μL) | 0.2 |
| dNTPmix(μL) | 3 |
| thermo pol Buffer(μL) | 3 |
| dH2O(μL) | 19.8 |
| TOTAL(μL) | 30 |
->95°C 5 min ->(95°C 1 min 50°C 30sec 72°C 1 min)×25 ->72°C 10 min
Digestion Test
[http://partsregistry.org/Part:BBa_I6206 BBa_I6206]
[http://partsregistry.org/Part:BBa_I6101 BBa_I6101]
[http://partsregistry.org/Part:BBa_I6042 BBa_I6042]
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462]
| Sample No | each |
| DNA(μL) | 2 |
| EcoRI(μL) | 0.2 |
| EcoRI Buffer(μL) | 1 |
| dH2O(μL) | 7 |
| TOTAL(μL) | 30 |
->30 min 37°C
Gel check
[http://partsregistry.org/Part:BBa_I6206 BBa_I6206](digestion product)
[http://partsregistry.org/Part:BBa_I6101 BBa_I6101](digestion product)
[http://partsregistry.org/Part:BBa_I6042 BBa_I6042](digestion product)
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462](digestion product)
[http://partsregistry.org/Part:BBa_I0642 BBa_I0642](natural)
| Sample No | each(digestion) | I0462(natural) |
| DNA(μL) | 10 | 1 |
| Loading Dye(μL) | 2 | 1 |
| dH2O(μL) | - | 4 |
| TOTAL(μL) | 12 | 6 |
Digestion
[http://partsregistry.org/Part:BBa_I0462 BBa_I0462]
| DNA(μL) | 25 |
| XbaI(μL) | 1 |
| PstI | 1 |
| Buffer 3(μL) | 4 |
| BSA(μL) | 4 |
| dH2O(μL) | 5 |
| TOTAL(μL) | 40 |
->3 hours 37°C
->Zymo ->10μL
Ligation
Vector:[http://partsregistry.org/Part:BBa_I7100 BBa_I7100]
Insert:[http://partsregistry.org/Part:BBa_R0062 BBa_R0062]
Insert:[http://partsregistry.org/Part:BBa_I0462 BBa_I0462]
| I7100 | 1.3 | 1.3 |
| R0062 | 2 | - |
| I0462 | 3.3 | - |
| Ligase(μL) | 1 | 1 |
| Ligase Buffer(μL) | 2 | 2 |
| dH2O(μL) | 0.4 | 5 |
| TOTAL(μL) | 10 | 10 |
->3 hours 37°C
->Transformation
