User:Nader A. Khamis/Notebook/Experimental Biological Chemistry AU Fall 2011/2011/09/20
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Forward Primer: 5' - CGA CGA TGA CGA TAA GCG ATG GGG ATC CGA ATT CGC - 3'
Reverse Primer: 5' - GCG AAT TCG GAT CCC CAT CGC TTA TCG TCA TCG TCG - 3'
Melting Temperature of Theoratical Primers (Tm) = 79.1°C Percent of GC content= 56%.
Forward primer used was:5' GAC GAT GAC GAT AAG GAT CGA TGG GGA TCC GAA 3'
Reverse primer used was: 5' TTC GGA TCC CCA CCA TCG ATC CTT ATC GTC ATC GTC 3'
Melting Temperature of actual primers = 64.6°C Percent of GC content = 51.5%
5 uL 10x reaction buffer
42 uL dH2O
1 uL dsDNA template (50 ng/uL)
1.25 uL Primer 1
1.25 uL Primer 2
1 uL dNTP mix (10 mM)
The final cycle was: 4°C for 24 hours.
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