Team:Chiba/Notebook/Calendar/29 September 2009

From 2009.igem.org

(Difference between revisions)
(→PCR)
(→Digestion Test)
 
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*Sample
*Sample
Vector(plux-GFP)
Vector(plux-GFP)
 +
 +
Insert(sfGFP)
 +
 +
 +
*Vector
 +
<table width="100" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
<td width="50">Vector DNA</td>
 +
<td width="50">30 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nco1</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BamH1</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer3</td>
 +
<td>5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BSA</td>
 +
<td>5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>NFW</td>
 +
<td>8 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Total</td>
 +
<td>50 &mu;L</td>
 +
</tr>
 +
</table>
 +
 +
 +
 +
*Insert
 +
<table width="100" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
<td width="50">Insert DNA</td>
 +
<td width="50">20 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Nco1</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BamH1</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer3</td>
 +
<td>3 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>BSA</td>
 +
<td>3 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>NFW</td>
 +
<td>2 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Total</td>
 +
<td>30 &mu;L</td>
 +
</tr>
 +
</table>
 +
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Took a picture
Took a picture
-
 
-
 
-
 
== Transformation ==
== Transformation ==

Latest revision as of 08:00, 3 October 2009

>Go to the Notebook page

(28_September_2009 <|>30_September_2009)



Contents

Gel electrophoresis

Yesterday's oparation is here.


  • Today's operation

10:40-

Mini Prep.


13:30-

Gel electrophoresis(135 V, 27 min)


---DNA Clean & Concentrator---


15:50-

Gel electrophoresis again(135 V, 27 min)

Digestion Test

  • Sample

Vector(plux-GFP)

Insert(sfGFP)


  • Vector
Vector DNA 30 μL
Nco1 1 μL
BamH1 1 μL
Buffer3 5 μL
BSA 5 μL
NFW 8 μL
Total 50 μL


  • Insert
Insert DNA 20 μL
Nco1 1 μL
BamH1 1 μL
Buffer3 3 μL
BSA 3 μL
NFW 2 μL
Total 30 μL


18:55-

Gel electrophoresis(135 V, 25 min)


19:45

Took a picture

Transformation

  • Sample

Competent Cells : XL10G(K)

Plasmid : plux-GFP(Cm)


21:45-

CmのプレートにまいてCultured it at 37 degrees Celsius.



Digestion

  • Sample

Vector : plux-GFP(p15A)

Insert : sfGFP

Enzymes : Nco1, BamH1


21:17

We added Enzymes and took it 37 degrees Celsius.


---3 h---


We took it 65 degrees Celsius for 酵素を失活させる


---20 min---

24:42

We kept it in refrigerator.



PCR

  • Template

plux-HSUTK


  • Element of mixtures
Template 1 μL
Primer(F) 2.5 μL
Primer(R) 2.5 μL
Buffer(x 10) 5 μL
dNTP 4 μL
DNA pol. 1 μL
dH2O 34 μL
Total 50 μL
Retrieved from "http://2009.igem.org/Team:Chiba/Notebook/Calendar/29_September_2009"