for
30 sec
Annealing/
Extension
68 °C for
3–5 min
68 °C for
6–10 min
Final extension
(optional)
68 °C for
3 min
(May reduce
background)
68 °C for
10 min
Hold 4 °C 4 °C
65 °C 20 sec
Extension 68 °C 20 min
For cycle 16-30:
Denaturation 94 °C 5-15 sec
Annealing 65 °C 20 sec
Extension 68 °C 20 min+15
sec/cycle
Final extension: 68 °C
results (see Table 1 and Figure 2).
■ This product is recommended for use in initial starting volumes of 100–500
microliters, but is limited to the volume capacity of the 96-well receiver
plate. For samples
fragment
Initial denaturation 96 °C 30 sec
For cycles 1-30:
Denaturation 94 °C 5-15 sec
Annealing 62-65 °C 30 sec
Extension 68 °C 20 –25 min
Final extension: 68 °C 30 min
Hold 4 °C
5. Evaluate
SURGXFWV�IURP� �FDVHLQ
(1 cm light path).2
FINAL ASSAY CONCENTRATION:
In a 4 ml reaction mix, the final concentrations are 45 mM Tris, 68 mM sodium chloride,
2 units SODVPLQRJHQ���������Z�Y�� �FDVHLQ
synthesis of fiscalin A. J. Org.
Chem., 68(2), 545-563 (2003).
10. Gliniski, J., et al., Surface properties of aqueous
solutions of L-leucine. Biophys. Chem., 84(2),
99-103 (2000).
GCY/RXR 8/
synthesis of fiscalin A. J. Org.
Chem., 68(2), 545-563 (2003).
10. Gliniski, J., et al., Surface properties of aqueous
solutions of L-leucine. Biophys. Chem., 84(2),
99-103 (2000).
GCY/RXR 3/07
seconds
annealing 60 °C, 30 seconds
extension 68 °C, 2 minutes 30 seconds
Final extension: 68 °C, 10 minutes
Soak: 4 °C
Note: If the assembled template is >2 kb, it may be
necessary to modify the PCR
seconds
annealing 60 °C, 30 seconds
extension 68 °C, 2 minutes 30 seconds
Final extension: 68 °C, 10 minutes
Soak: 4 °C
Note: If the assembled template is >2 kb, it may be
necessary to modify the PCR
fragment
Initial denaturation 96 °C 30 sec
For cycle 1-30:
Denaturation 94 °C 5-15 sec
Annealing 62-65 °C 30 sec
Extension 68 °C 20 -25 min
Final extension 68 °C 30 min
Hold 4 °C
5. Evaluate