PCR reaction mix, 8 tubes, 50 μL each: Mix contains Taq DNA polymerase, MgCl2, and dNTP's
DNA / primer mix, 8 tubes, 50 μL each: Each mix contains a different template DNA. All tubes have the same forward and reverse primer.
Strip of empty PCR tubes
Disposable pipette tubes
Cup for discarded tips
Micropipettor
OpenPCR machine
PCR Reaction Sample List
Tube Label
PCR Reaction Sample
Patient ID
G15 +
Positive control
66684 (patient 1)
G15 -
Negative control
40117 (patient 2)
G15 1-1
Patient 1, replicate 1
G15 1-2
Patient 1, replicate 2
G15 1-3
Patient 1, replicate 3
G15 2-1
Patient 2, replicate 1
G15 2-2
Patient 2, replicate 2
G15 2-3
Patient 2, replicate 3
DNA Sample Set-up Procedure
Step 1
Step 2
Step 3...
OpenPCR program
HEATING LID: 100 °C
INITIAL STEP: 95 °C for 2 minutes
NUMBER OF CYCLES: 35
Denatures at 95 °C for 30 seconds, Anneal at 57 °C for 30 seconds, and Extend at 37 °C for 30 seconds
FINAL STEP: 72 °C for 2 minutes
FINAL HOLD: 4 °C
Research and Development
PCR - The Underlying Technology
The PCR Reaction
The four components necessary in a PCR reaction are the template DNA, the primers, taq polymerase, and dNTP's (deoxyribonucleotides). The template DNA is the DNA from the subject that will be duplicated in the PCR reaction. The primers are short pieces of DNA that are made in the lab which will attach to the segments of DNA that are to be copied. The taq polymerase is a naturally occurring protein that attaches to the primers and begins adding nucleotides to the DNA. The dNTP's are A's, C's, G's, and T's that are needed to build the DNA.
Q2 During the initial step the temperature is held at 95°C for 2 minutes to activate the heat activated polymerases. During the Denature step the solution is heated to 95°C for 30 seconds to denature the DNA. Where the bonds between the complementary bases of the two DNA strands is disrupted allowing the two DNA strands to separate. During the Anneal step the solution is cooled from 95°C to 57°C for 30 seconds
BME 100 Fall 2014
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