Engineering BioBrick vectors from BioBrick parts/Colony PCR protocol

Solutions/reagents: PCR SuperMix High Fidelity VF2 primer  (5'-TGCCACCTGACGTCTAAGAA-3')  VR primer  (5'-ATTACCGCCTTTGAGTGAGC-3')  colony suspension  (1 colony diluted in 100 ?l water) de-ionized water</ul> Parameters: X - concentration of primers (µmole/µl)</li></ul> Equipment: Thermocycler</li>Sterile 0.6-ml tubes</li></ul> Steps: <ol>  PCR mix  Use the following table as a checklist for preparing the reaction in sterile 0.6-ml microcentrifuge tube (1): </li>   PCR conditions  Program a standard thermocycler to run the reaction using the following parameters: Initial denaturation <ul>Denature: <font color=#357EC7>95°C , <font color=#357EC7>15 mins </li></ul>Thermocycling <ul>No. of cycles: <font color=#357EC7>39 </li>Denature: <font color=#357EC7>94°C , <font color=#357EC7>30 secs </li>  Anneal: <font color=#357EC7>62°C , <font color=#357EC7>30 secs </li> Elongate: <font color=#357EC7>68°C , <font color=#357EC7>3.5 mins </li></ul>Termination<ul>Elongate: <font color=#357EC7>68°C , <font color=#357EC7>20 mins </li>Hold: <font color=#357EC7>4°C , until removed from machine </li></ul></li> </ol> TOTAL TIME REQUIRED FOR THE COMPLETION OF THE PROTOCOL :<font color=#357EC7>~ 3 hrs, 35 mins