First step PCR protocol						
						
	Per Rxn	26		94¡C	3:00	
Master Mix (2x)	12.5	325		94¡C	0:45	
Water	7.5	195		50¡C	1:00	35x
Primer F (10 uM)	0.5	13		72¡C	1:30	
Primer R (10 uM)	0.5	13		72¡C	10:00	
DNA	4					
Total Volume	25	520				
						
						
Second step PCR protol						
						
	Per Rxn	26		94¡C	3:00	
Master Mix (2x)*	12.5	325		94¡C	0:30	
Water	5	130		55¡C	0:30	8x
PrimerF&R	5			72¡C	0:45	
DNA	2.5			72¡C	7:00	
Total Volume	25	455