BME100 s2016:Group8 W1030AM L5

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.png BME 100 Spring 2016 Home
People
Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
Course Logistics For Instructors
Photos
Wiki Editing Help
BME494 Asu logo.png

OUR TEAM

Name: Abdulrahman Hassan M Alamri
Name: Eric Barrientos
Name: Ethan Blank
Name: Melanie Parke
Name: Vanessa Sanders
Name: Erin Sussex


LAB 5 WRITE-UP

PCR Reaction Report

Members of our team had extensive previous experience pipetting which led to little difficulty in setting up the reaction. The pre-lab reading was a good review of the technique but was aimed at people with less experience with the process. Because we knew the difference between the first and second stops our measurements were as accurate as possible, but there is always a possibility of error in a human-driven procedure. In a couple of the tubes there was a small amount of sample left. This could have been due to taking air into the pipettor instead of liquid or an accidental adjustment of the volume setting on the handle of the pipettor. In the final part of the set up, we came up with a good labeling scheme, but we had to relabel the sides of the tubes before putting them into the PCR machine in case the intense heat removed the ink from the lids.

Fluorimeter Procedure

Web camera set-up

This experiment used an iPhone 6 camera without flash. None of the other settings could be changed manually. The phone was placed in the cradle so that the lens was level with and at a right angle to the slide and 8 cm from the front of the slide for each photograph and focused.


Placing Samples onto the Fluorimeter

  1. Place a new slide onto the fluorimeter with the hydrophobic side facing up.
  2. Pipet 80 microliters of the SYBR Green onto the first two rows of the slide in a single drop. Using a new pipet tip, add 80 microliters of the sample to the drop.
  3. Using the switch on the side of the fluorimeter, turn on the LED light.
  4. Set up the phone camera as described above.
  5. Move the box around the fluorimeter to block as much ambient light as possible.
  6. Take 3 photographs of the drop.
  7. Pipet the drop off of the slide and dispose of the sample.
  8. Move the slide back 2 rows so that the next two rows are illuminated by the LED light and turn off the light.
  9. Repeat steps 2-8 for each remaining sample replacing the slide once all rows have been used.


Data Collection and Analysis

Images of High, Low, and Zero Calf Thymus DNA


Calibrator Mean Values

Description of image


Calibration curves

Description of image

Images of Our PCR Negative and Positive Controls


PCR Results: PCR concentrations solved

Description of image



PCR Results: Summary

  • Our positive control PCR result was ____ μg/mL
  • Our negative control PCR result was ____ μg/mL


Observed results

  • Patient _____ :
  • Patient _____ :


Conclusions

  • Patient _____ :
  • Patient _____ :