BME100 s2017:Group9 W8AM L5

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Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
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OUR TEAM

Name: Emma Spencer
Name: Aswathy Lawrence
Name: Jocelyn Alvar
Name: Brett Nasch
Name: Anna Seminara
Name: Will Anderson


LAB 5 WRITE-UP

PCR Reaction Report

Fluorimeter Procedure

Imaging set-up



Placing Samples onto the Fluorimeter

  1. 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
  2. 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).
  3. 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.
  4. Place the now labeled tubes in a rack.
  5. 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)
  6. 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.
  7. Repeat steps 5 and 6 for the rest of the labeled tubes with the proper DNA mix/primer for each one.
  8. Ensure each lid is tightly shut.
  9. 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

Name: High 5 microliter
Name: Low .5 microliter
Name: Low and Image J
Name: Zero microliter


Calibrator Mean Values


Name: Calibration mean Table


Calibration curves

Calibration Curve 1
Curve 2

Images of Our PCR Negative and Positive Controls


PCR Results: PCR concentrations solved

PCR Solutions table
PCR Positive Control
PCR Negative Control


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.