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Category: Experimental Procedures
Author: Admin eLabJournal

Labels: Cloning, DNA

Materials
 

- PCR reaction buffer
- dNTPs (10X)
- MgCl2  ( mM )
- DMSO (100%)
-  polymerase  (  U µl-1)
- Template
- Primers
- PCR machine

Experiment settings

- Number of samples:    
- Reaction volume: µl  make % more
- Template Type:    
- Template Name:   µl /reaction
- Primer 1:   µl /reaction
- Primer 2:   µl /reaction

Step 1
-Prepare a the following reaction mix:

- PCR reaction buffer (10X) µl 
- dNTP mix (10X) µl 
- mM MgCl2  = µl 
- % DMSO = µl 
-  polymerase (  U µl-1) µl 
- Template:     µl 
- dH20 µl 
Total µl 


Step 2
Vortex the reaction mixture and add µl  to   PCR tubes
Step 3
Add primers to each tube:
µl 
  
µl   
Step 4
Mix the tubes, spin down briefly and program the PCR machine to start the reaction. After PCR proceed to DNA agarose gel electrophoresis

Keep the reaction mix on ice as much as possible, especially after the polymerase enzyme is added!