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Team:METU/test/Protocols
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<p>Prepare mastermix according to your sample amount ( except Taq and template)<p> | <p>Prepare mastermix according to your sample amount ( except Taq and template)<p> | ||
1X | 1X | ||
| - | dNTP 1 µL | + | <ul> |
| - | Buffer 5 µL | + | <li>dNTP 1 µL</li> |
| - | MgCl2 3 µL | + | <li>Buffer 5 µL</li> |
| - | **Forward Primer 0,5 µL | + | <li>MgCl2 3 µL</li> |
| - | **Reverse Primer 0,5 µL | + | <li>**Forward Primer 0,5 µL</li> |
| - | Template - | + | <li>**Reverse Primer 0,5 µL</li> |
| - | Taq Polymerase 0,2 µL | + | <li>Template - </li> |
| - | dH2O 39,8 µL | + | <li>Taq Polymerase 0,2 µL</li> |
| - | TOTAL 50 µL | + | <li>dH2O 39,8 µL </li> |
| + | <li>TOTAL 50 µL</li> | ||
<p> | <p> | ||
* You should add ingredients from largest amount to smallest amount. | * You should add ingredients from largest amount to smallest amount. | ||
Revision as of 11:23, 23 September 2012
Protocols[gizle] |
COLONY PCR:
Prepare mastermix according to your sample amount ( except Taq and template)
1X
- dNTP 1 µL
- Buffer 5 µL
- MgCl2 3 µL
- **Forward Primer 0,5 µL
- **Reverse Primer 0,5 µL
- Template -
- Taq Polymerase 0,2 µL
- dH2O 39,8 µL
- TOTAL 50 µL
* You should add ingredients from largest amount to smallest amount. * Before addition of primers and template you can do vortex. ** First you should pour ddH20 onto dried primers according to the amount written in the primer sheet then you should dilute (1:10 ) it in to new eppendorf(10 ml primer+ 90 ml ddH20)
Pour your samples into PCR tubes and add template that you pick from the petri with toothpick or tip and finally add taq polymerase. Then place your samples into the PCR machine and do regular PCR.
