CDC Ad Hexon Gene PCR Master Mix Worksheet and Thermal Cycler Program
Date: ___________
Added
TOTAL
1 Rxn
xn
Initials: ________________
= n Rxn
Lot #
37.2 μl
dH20 (Gibco Ultra Pure)
5.0 μl
Invitrogen 10X PCR buffer minus Mg
0.4 μl
dNTP (25mM)
1.5 μl
50 mM MgCl2
0.20 μl
Ben (date:___________)Forward Primer (50uM)
0.20 μl
Marcy (date:_________)Reverse Primer (50uM)
0.50 μl
Invitrogen Platinum Taq DNA Polymerase
(5 U/µL)
45.0 μl
Exp.
TOTAL:
5.0 μl
TOTAL
Reagent
Qiagen Treated Sample
50.0μl
Preparation of Master Mix:
- Gather reagents and samples and keep on ice, check that all reagent
concentrations are correct prior to pipetting
- Add all reagents, except the treated sample, in the order as listed above
- Vortex mixture and centrifuge briefly
- Aliquot 45.0 μl of the master mix into micro amp tubes
- Add 5.0 μl of the respective samples to the micro amp tubes, mix well and
centrifuge briefly
 Always include a water negative control in each run
- Continue with the PCR Program on the Whatman Biometra Thermal
Cycler
PCR Program for CDC Adeno Hexon gene:
- Program #19: CDC AD HEX
 Double check to see if the program has not changed
 Lid Temperature: 104°
 Program Time: 1 hours 41 minutes 20 seconds
Step
1
2
3
4
5
6
Temp
94°C
94°C
50°C
72°C
72°C
4°C
Time
2 min
30 sec
30 sec
1 min
5 min
Pause
←
#
2
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