Table 2.

PCRs and thermal-cycling parameters for amplification ofCryptosporidium DNA

PCR method (reference)MgCl2 concn (mM)Amt of Taq(U)Cycle conditionsNo. of cyclesRestriction site(s)
Initial denaturationDenaturationAnnealingExtensionFinal extension
Laxer (15)2195°C, 10 min94°C, 1 min56°C, 90 s72°C, 90 s72°C, 7 min45
Laberge (14)2195°C, 10 min94°C, 1 min56°C, 90 s72°C, 90 s72°C, 7 min45
Wagner-Wiening (28)2294°C, 5 min94°C, 1 min50°C, 1 min72°C, 1 min72°C, 10 min35 BamHI
Morgan (18)2195°C, 10 min94°C, 1 min58°C, 90 s72°C, 90 s72°C, 10 min45
Johnson (13)4194°C, 5 min94°C, 30 s55°C, 30 s72°C, 1 min72°C, 10 min40
Awad-El-Kariem (1)2294°C, 5 min94°C, 90 s47°C, 90 s72°C, 3 min72°C, 10 min45 MaeI
Rochelle (24)2194°C, 2 min94°C, 30 s55°C, 30 s72°C, 90 s72°C, 5 min40
Bonnin (3)2.51.594°C, 20 min94°C, 90 s45°C, 90 s72°C, 90 s72°C, 10 min45 HinfI, RsaI