BME100 s2017:Group3 W1030AM L5: Difference between revisions

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'''Placing Samples onto the Fluorimeter'''
'''Placing Samples onto the Fluorimeter'''
<!-- INSTRUCTIONS: In the space below, in your own words write the steps you performed to place samples onto the fluorimeter -->
<!-- INSTRUCTIONS: In the space below, in your own words write the steps you performed to place samples onto the fluorimeter -->
# ''[Instructions: Step one, in your own words]''
 
# ''[Instructions: Step two, in your own words]''
# Collect and label all necessary samples.
# ''[Instructions: Step three, in your own words]''
# Add 80 microliters of SYBR Green to the rougher side of the glass slide. It will appear as a bubble.
# ''[Instructions: Step etc., in your own words]''
# Discard the pipette tip and replace it.
# Add 80 microliters of one of the Calf Thyme samples to the bubble of SYBR Green.
# Adjust the glass slide so that the blue LED light shines through the bubble made in step 1 and 2 so that it hits the black fiber optic fitting.
 


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Revision as of 12:30, 29 March 2017

BME 100 Spring 2017 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

OUR TEAM

Name: Caden Keller
Name: Karolena Lein
Name: Claudia Fragoso
Name: Devin Dulay
Name: Abigail Call
Name: Madison Ott


LAB 5 WRITE-UP

PCR Reaction Report

Using new equipment for the first time was difficult but the pre-lab gave easy to understand instructions. It took some time for us to understand the difference between the first and second stop but by looking at the directions we were able to figure it out. The first stop takes in the liquid and the second stop releases it. Between the two stops we could feel a click within the pipette. Once we were done mixing the controls and patients tests there was a trace amount of DNA samples and PCR reaction mix left within their tubes. Because there was small varying amounts of liquid left of the DNA samples and PCR reaction mix the final reactions did not have exactly the same amount of liquid. With more practice using the pipette it would be easier to leave less amount of DNA samples and PCR reaction mix and have the same amount of liquid in the final reactions. During the experiment we did not have to change our labeling scheme, we used the same scheme that we planned to use.

Fluorimeter Procedure

Imaging set-up
The first step was placing the slide onto the fluorimeter and the phone onto the cradle to hold it. Then the height of the flourimeter and the phone was adjusted so that the phone would be able to just see above the edge of the slide. 80 microliters of the Green 1 solution was added to the first section of the slide, followed by 80 microliters of the calibration solution. The slide was then adjusted so that the light from the fluorimeter was shining on the center of the 160 microliter solution. The camera was adjusted again and the distance from the fluorimeter was recorded. The lightbox was placed over the setup, and the camera was set on a timer. The camera was activated and the final flap was lowered three times to get three photos of the solution. The slide was removed and the process repeated with a new slide and new calibration solutions.



Placing Samples onto the Fluorimeter

  1. Collect and label all necessary samples.
  2. Add 80 microliters of SYBR Green to the rougher side of the glass slide. It will appear as a bubble.
  3. Discard the pipette tip and replace it.
  4. Add 80 microliters of one of the Calf Thyme samples to the bubble of SYBR Green.
  5. Adjust the glass slide so that the blue LED light shines through the bubble made in step 1 and 2 so that it hits the black fiber optic fitting.



Data Collection and Analysis

Images of High, Low, and Zero Calf Thymus DNA


Calibrator Mean Values


TABLE GOES HERE


Calibration curves


Images of Our PCR Negative and Positive Controls


PCR Results: PCR concentrations solved

TABLE GOES HERE


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 _____ :


Gel Electrophoresis