the next page. For maximum yield and specificity, temperatures
and cycling times should be optimized for each new template target
or primer pair.
7. Place the PCR tubes in the thermal cycler and start the cycling
Note: After amplification, samples can be stored overnight at 28C
or at 20C for longer storage.
PCR Protocol Using HotStarTaq Master Mix
This protocol serves only as a guideline for PCR amplification. Optimal
reaction conditions, such as incubation times and temperatures, and
amount of template DNA, may vary and need to be determined individually.
Each PCR program should be started with an initial activation
step of 15 min at 95C to activate HotStarTaq DNA Polymerase (see
step 6 of this protocol).
HotStarTaq Master Mix provides a final concentration of 1.5
mM MgCl2 in the final reaction mix, which will produce satisfactory
results in most cases.
However, if a higher Mg2
+ concentration is required, prepare a stock
solution containing 25 mM MgCl2.
Set up reaction mixtures in an area separate from that used
for DNA preparation or PCR product analysis.
Use disposable tips containing hydrophobic filters to minimize
1. Thaw primer solutions.
Mix well before use.
prepare a primer mix of an appropriate concentration
(see Table 4) using the water provided.<
Source:Page: All 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 Related biology technology :1
. HotStarTaq DNA Polymerase2
. QIAGEN Multiplex PCR Handbook3
. TransMessenger Transfection Reagent Handbook4
. Transfection Reagent Selector Kit Handbook