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Category: Experimental Procedures
Author: Admin eLabJournal

Labels: DNA, Cloning

Materials
 

- PCR reaction buffer (10X)
- 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 MgCl= µl 
- DMSO  = µl 
-  polymerase (  U µl-1) µl 
- Primer 1:   µl 
- Primer 2:   µl 
- dH20 µl 
Total µl 


Step 2
Vortex the reaction mixture and add µl  to   PCR tubes
Step 3
Add template to each tube:
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!