User:Klare Lazor/Notebook/Chem-496-001/2011/09/20
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Objectivemutation of GFP to contain a cysteine residue just after the enterokinase clevage site through PCR. DescriptionQuick Change Manual was used in order to determine the forward and reverse PC primers. The PCR reaction was performed by using previously prepared primers. 43uL of ddH2 was added. 1uL on .5u/uL of template DNA, 1uL of a 10mM dNTP solution, and 1.25uL of a 100ng/uL solution of forward and reverse primers was pipeted into the solution. 1uL of enzymes was added to the solution, along with 25uL of wax to prevent evaporation, before placed in the thermocylcer. DataPrimers: FORWARD: 5' GAC GAT GAC GAT AAG GAT CGA TGG GGA TCC GAA 3' REVERSE 5' TTC GGA TCC CCA CCA TCG ATC CTT ATC GTC ATC GTC 3' [edit] NotesPCR Background
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