LAB 5 WRITE-UP
PCR Reaction Report
The team experienced no trouble with pipetting the samples for Lab C. Although there were various skill levels in the group regarding micro-pipetting prior to Lab C, the pre-lab reading assisted in helping those that had not previously worked with micropipettes. During the lab and all proceeding labs, we thought it best to utilize two micropipettors: one experienced and one using a micropipette for the first time. With this setup, we always had one knowledgeable member paired and prepared to assist the less experienced member while they improve their skills.
The final reactions did end up having the exact same amount of liquid due to the spot on precision of the micro-pipettes. There was some liquid left in the tubes containing DNA samples, however the PCR reaction mix was used entirely by the end of the lab. Our labeling scheme did not have to be changed after the initial labeling.
Smart Phone Camera Settings
- Type of Smartphone: iPhone 6
- Flash: Off
- ISO setting: 2000
- White Balance: Auto White Balance
- Exposure: 1/15 sec
- Saturation: N/A
- Contrast: N/A
*Note: All setting specifics obtained through image metadata via OpenWetWare.
- Distance between the smart phone cradle and drop = 11.00 cm
Placing Samples onto the Fluorimeter
- Slide glass slide into designated grooves on the fluorimeter
- After obtaining the necessary quantity of dye solution in the micropipette, start by placing a single drop in the middle circular groove of the first row on the glass slide. Continue by placing another drop in the next groove back. Place a third drop in-between the two preexisting drops and continue dispensing the dye in the pipette until there is a globular collection of dye in the middle of the slide.
- Repeat Step #2 with the necessary quantity of DNA solution.
- Center the DNA/Dye solution bubble so that it is cut in half by the light of the fluorimeter.
Data Collection and Analysis
Images of High, Low, and Zero Calf Thymus DNA
"High Concentration (5micrograms/mL sample):"
"Low Concentration (.5micrograms/mL sample):"
"No Concentration (0micrograms/mL sample):"
Calibrator Mean Values
Images of Our PCR Negative and Positive Controls
PCR Results: PCR concentrations solved
PCR Results: Summary
- Our positive control PCR result was 26.73 μg/mL
- Our negative control PCR result was -4.76 μg/mL
- Patient 51283 : The images for Patient 51283 were most similar to the images taken of the negative control. -57.01 μg/mL
- Patient 60999 : The images for Patient 60999 were most similar to the images taken of the positive control. 23.23 μg/mL
- Patient 51283 : This patient is negative for the tested SNP as the PCR Product Concentration is nearest to that of the negative control.
- Patient 60999 : This patient is positive for the tested SNP as the PCR Product Concentration is nearest to that of the positive control.