Materials Needed: lab coat and disposable gloves; PCR reaction mix, 8 tubes, 50 μL each; DNA/ primer mix, 8 tubes, 50 μL each; a strip of empty PCR tubes; disposable pipette tips; cup for discarded tips; micropipettor; OpenPCR machine
To enable all of the tubes to fit into the OpenPCR machine, cut the empty strip of PCR tubes in half (two strips of 4 linked tubes).
Take the empty tubes and label them with the Tube Labels with a black marker. Take special care to label on the sides of the tubes, and not the lids.
Place the now labeled tubes in a rack.
Starting with the positive control labeled empty tube, transfer 50 μL of PCR reaction mix into the tube using proper pipetting technique (which includes discarding the disposable tip into the collection cup to not be used again)
With an unused pipette tip, transfer the positive control DNA/ primer mix into the same, "positive control" tube. The total volume in your positive control PCR reaction tube should be 100 μL.
Repeat steps 5 and 6 for the rest of the labeled tubes with the proper DNA mix/primer for each one.
Ensure each lid is tightly shut.
Place tubes into assigned PCR machine along with another 8 tubes to fill all 16 slots in the heating block.
Data Collection and Analysis
Images of High, Low, and Zero Calf Thymus DNA
Calibrator Mean Values
Calibration curves
Images of Our PCR Negative and Positive Controls
PCR Results: PCR concentrations solved
PCR Results: Summary
Our positive control PCR result was 0.002156168178 μg/mL
Our negative control PCR result was 0.0002380148 μg/mL
Observed results
Patient 37823 : Lightly green - positive 0.002156168178 μg/mL
Patient 49958 : Lightly green - positive 0.0002380148 μg/mL
Conclusions
Patient 1 positive : qualitatively the images appeared to looked closer to the positive control than the negative control and the numbers were larger than those of the negative control. We believe our initial values for the positive control were off through some error in setup or calculation.
Patient 2 positive : qualitatively the images appeared to looked closer to the positive control than the negative control and the numbers were larger than those of the negative control. We believe our initial values for the positive control were off through some error in setup or calculation.