Team:Chiba/Notebook/Calendar/29 September 2009

From 2009.igem.org

(Difference between revisions)
(→Gel electrophoresis)
(→PCR)
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*Template
*Template
plux-HSUTK
plux-HSUTK
 +
 +
 +
*Element of mixtures
 +
 +
<table width="200" border="1" cellpadding="0"  cellspacing="0" bordercolor="#000000"><tr>
 +
<td width="100">Template</td>
 +
<td width="100">1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Primer(F)</td>
 +
<td>2.5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Primer(R)</td>
 +
<td>2.5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer(x 10)</td>
 +
<td>5 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>dNTP</td>
 +
<td>4 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>DNA pol.</td>
 +
<td>1 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
 +
<td>34 &mu;L</td>
 +
</tr>
 +
<tr>
 +
<td>Total</td>
 +
<td>50 &mu;L</td>
 +
</tr>
 +
</table>

Revision as of 07:50, 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)


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
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