"
Team:UNIPV-Pavia/Protocols/Pcr
From 2008.igem.org
(Difference between revisions)
(New page: {| style="color:#000000;background-color:#ffffff;" cellpadding="3" cellspacing="1" border="3" bordercolor="#3128" width="80%" align="center" !align="center"|30px [[Team...) |
|||
| Line 41: | Line 41: | ||
*'''VR primer''' | *'''VR primer''' | ||
<br> | <br> | ||
| - | *For every DNA sample you want to amplify: | + | *For every DNA sample you want to amplify, put: |
| - | **2 | + | **2 µl buffer |
| + | **0.6 µl MgCl2 | ||
| + | **0.4 µl dNTPs | ||
| + | **1 µl DNA (or ddH2O for blank sample) | ||
| + | **0.2 µl Taq Polymerase | ||
| + | **250 nM VF2 primer | ||
| + | **250 nM VR primer | ||
| + | **A proper amount of ddH2O to have 20 µl of total reaction volume | ||
| + | *into a | ||
<br> | <br> | ||
Revision as of 14:31, 11 June 2008
 Home
|  The Team
|  The Project
|  Biological Safety
|  Parts Submitted to the Registry
|
|---|---|---|---|---|
 Dry Lab
|  Wet Lab
|  Modeling
|  Protocols
|  Activity Notebook
|
The protocols we used
- LB medium preparation
- Plasmid resuspension from IGEM paper spots
- Transformation
- Plasmid extraction
- BioBrick digestion with restriction enzymes
- DNA gel extraction
- Antarctic Phosphatase
- Ligation
- PCR
Antarctic Phosphatase
(estimated time: 1 hour and 30 min)
Materials needed:
- MgCl2
- Buffer
- dNTPs
- ddH2O
- Taq Polymerase
- VF2 primer
- VR primer
- For every DNA sample you want to amplify, put:
- 2 µl buffer
- 0.6 µl MgCl2
- 0.4 µl dNTPs
- 1 µl DNA (or ddH2O for blank sample)
- 0.2 µl Taq Polymerase
- 250 nM VF2 primer
- 250 nM VR primer
- A proper amount of ddH2O to have 20 µl of total reaction volume
- into a
