PCR with Taq DNA Polymerase (NEB):

Using the guidelines of the provided Taq DNA Polymerase with Standard Taq Buffer (M0273) protocol.

1. Reaction Setup:
   
   2.5 µl 10X Standard Taq Reaction buffer
   2.5 µl dNTPS [2 mM]
   0.5 µl Forward Primer [10 µM]
   0.5 µl Reverse Primer [10 µM]
   1 µl Template DNA
   0.125 µl Taq DNA Polymerase
   17.875 µl MilliQ Water
   

   ---

   25 µl Total Reaction




2. PCR Program:
   

   Step	Temperature	Time
   Initial Denaturation	95 °C	30 seconds /5 Minutes for Colony PCR
   35 Cycles	95 °C	30 seconds
   	45 °C – 72 °C	60 seconds
   	68 °C	1 minute / 1 kb
   Final Extension	68 °C	5 minutes
   Hold	4 °C	forever