Polymerase Chain Reaction (PCR) protocol

Solutions/reagents: BufferdNTPsMgCl 2DMSOUpstream primerDownstream primerTemplate DNAdH2ODNA Polymerase</li></ul> Equipment: Thermocycler</li>Reaction tubes</li></ul> Steps: <ol>  For Promega GoTaq:  Set up a reaction in reaction tube (1) as follows on ice: </li>   For Roche High Fidelity:  Set up a reaction in reaction tube (1) as follows on ice:  </li>   An example program:  Program a standard thermocycler to run the reaction using the following parameters: Initial denaturation <ul>Denature: <font color=#357EC7>96°C , <font color=#357EC7>5 mins </li></ul><font color = "#800517">This denatures any double stranded DNA. Thermocycling <ul>No. of cycles: <font color=#357EC7>25 </li>Denature: <font color=#357EC7>96°C , <font color=#357EC7>1 min </li>  Anneal: <font color=#357EC7>55°C , <font color=#357EC7>30 secs </li> Elongate: <font color=#357EC7>72°C , <font color=#357EC7>1 min </li></ul>Termination<ul>Elongate: <font color=#357EC7>72°C , <font color=#357EC7>5 mins </li>Hold: <font color=#357EC7>4°C , until removed from machine </li></ul><font color = "#800517">Final extension for incomplete strands and holding of sample at low temperature. </li> </ol> TOTAL TIME REQUIRED FOR THE COMPLETION OF THE PROTOCOL :<font color=#357EC7>~ 1 hr, 15 mins