BME100 s2015:Group6 9amL5: Difference between revisions

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{| style="wikitable" width="300px"
{| style="wikitable" width="300px"
|- valign="top"
|- valign="top"
| [[Image:"setup3".jpg|100px|thumb|Name: Set Up]]  
| [[Image:"setup3".jpg|500px|thumb|Name: Set Up]]  
|}
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{| {{table}} width=700
{| {{table}} width=700
|-
|-
| [[Image:"Lab6water".jpg|100px|thumb|Name: Water]]   
| [[Image:"Lab6water".jpg|500px|thumb|Name: Water]]   
|-
|-
| [[Image:"Lab60.25".jpg|100px|thumb|Name: O.25]]  
| [[Image:"Lab60.25".jpg|500px|thumb|Name: O.25]]  
|-
|-
| [[Image:"Lab60.5".jpg|100px|thumb|Name: 0.5]]  
| [[Image:"Lab60.5".jpg|500px|thumb|Name: 0.5]]  
|-
|-
| [[Image:"Lab61".jpg|100px|thumb|Name: 1]]   
| [[Image:"Lab61".jpg|500px|thumb|Name: 1]]   
|-
|-
| [[Image:"Lab62".jpg|100px|thumb|Name: 2]]  
| [[Image:"Lab62".jpg|500px|thumb|Name: 2]]  
|-
|-
| [[Image:"Lab65".jpg|100px|thumb|Name: 5]]  
| [[Image:"Lab65".jpg|500px|thumb|Name: 5]]  
|}
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{|{{table}}
{|
| align="center" style="background:#f0f0f0;"|'''Concentration'''
| align="center" style="background:#f0f0f0;"|'''Concentration'''
| align="center" style="background:#f0f0f0;"|'''Area'''
| align="center" style="background:#f0f0f0;"|'''Area'''
| align="center" style="background:#f0f0f0;"|'''Internal Density'''
| align="center" style="background:#f0f0f0;"|'''Mean'''
|  
| align="center" style="background:#f0f0f0;"|'''RawIntDen Drop'''
| align="center" style="background:#f0f0f0;"|'''Raw IntDen Background'''
| align="center" style="background:#f0f0f0;"|'''Raw IntDen Drop- Background'''
|-
| 0||582||30||17609||2428||15181
|-
|-
| .25||1488||69984
| 0.25||582||39.7||23098||6099||16999
|-
|-
| .5||1456||73079
| 0.5||582||39.6||23071||4984||18087
|-
|-
| 1||1456||72004
| 1||582||26.321||15319||909||14410
|-
|-
| 2||1321||65740
| 2||582||126.3||73506||4821||68685
|-
|-
| 5||1264||60354
| 5||582||112.4||65415||2478||62937
|-
|-
|
|}
|}


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'''Calibration curve'''<br>
'''Calibration curve'''<br>
<!-- INSTRUCTIONS: Place an image of your Excel plot with a line of best fit here. -->
<!-- INSTRUCTIONS: Place an image of your Excel plot with a line of best fit here. -->
[[Image:Calibration6.png]]




'''PCR Results Summary'''
'''PCR Results Summary'''
<!-- INSTRUCTIONS: You completed 8 PCR reactions and used the SYBR Green I staining and imaging technique to measure the amount of amplified DNA in each PCR reaction. You used a standard curve (based on known concentrations of calf thymus DNA) to convert INTDEN values into DNA concentration. Your positive control and negative control samples should be used as '''threshold''' values for determining whether an unknown (patient) sample is truly positive or negative. Replace the underscore with your claculated initial concentration values.-->
<!-- INSTRUCTIONS: You completed 8 PCR reactions and used the SYBR Green I staining and imaging technique to measure the amount of amplified DNA in each PCR reaction. You used a standard curve (based on known concentrations of calf thymus DNA) to convert INTDEN values into DNA concentration. Your positive control and negative control samples should be used as '''threshold''' values for determining whether an unknown (patient) sample is truly positive or negative. Replace the underscore with your claculated initial concentration values.-->
* Our positive control PCR result was ____ μg/mL
* Our positive control PCR result was 5 μg/mL
* Our negative control PCR result was ____ μg/mL
* Our negative control PCR result was 0 μg/mL


<u>Observed results</u>
<u>Observed results</u>
<!-- INSTRUCTIONS: Replace the underscore with each patient ID. After the colon, write both a qualitative (what the images looked like) and a quantitative description (μg/mL) of what you observed -->
<!-- INSTRUCTIONS: Replace the underscore with each patient ID. After the colon, write both a qualitative (what the images looked like) and a quantitative description (μg/mL) of what you observed -->
* Patient _____ :  
* Patient 73039 : The sample did not contain the allele so it did not glow green. The sample was observed to be 0 μg/mL.   
* Patient _____ :
* Patient 40429 : The sample did contain the allele so it glowed green. The sample was observed to be 2 μg/mL.


<u>Conclusions</u>
<u>Conclusions</u>
<!-- INSTRUCTIONS: Compare each patient's results to the positive control value and the negative control value. Draw a final conclusion for each patient (positive or negative) and explain why you made that conclusion. -->
<!-- INSTRUCTIONS: Compare each patient's results to the positive control value and the negative control value. Draw a final conclusion for each patient (positive or negative) and explain why you made that conclusion. -->
* Patient _____ :
* Patient 73039 : This patient does not have coronary artery disease.
* Patient _____ :
* Patient 40429 : This patient does have coronary artery disease.
 
 


<br><br>
<br><br>
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<!-- INSTRUCTIONS: This content is from PCR Lab D. Write a summary, at least five sentences long, about the disease SNP in your own words. -->
<!-- INSTRUCTIONS: This content is from PCR Lab D. Write a summary, at least five sentences long, about the disease SNP in your own words. -->


The disease SNP is associated with lipoprotein lipase, which helps regulate phospholipase activity. It caused by a genetic mutation in the 8th chromosome. More specifically, in the 5' strand, changing AAT to AGT. As a result, it causes coronary artery disease in homo sapiens. Coronary artery disease, or heart disease, is one of the deadliest diseases in America.


'''Primer Design and Testing'''
'''Primer Design and Testing'''
<!-- INSTRUCTIONS: Write a short summary of the results of your primer test. Underneath your summary, include a screen capture of the results web page. You may crop the image so that it only includes the relevant information. -->
<!-- INSTRUCTIONS: Write a short summary of the results of your primer test. Underneath your summary, include a screen capture of the results web page. You may crop the image so that it only includes the relevant information. -->


At the end of the primer test, the non-diseased primer test gave a 220bp reading, while the diseased primer had no match.




{| style="wikitable" width="700px"
|- valign="top"
| [[Image:"nondiseased".jpg|500px|thumb|Name: Nondieased]]
| [[Image:"diseased".jpg|500px|thumb|Name: Diseased]]
|}




<!-- Do not edit below this line -->
<!-- Do not edit below this line -->
|}
|}

Latest revision as of 12:41, 1 April 2015

BME 100 Spring 2015 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: Sara Belko
Name: Luc Tieu
Name: Priscilla Delgado
Name: Al Imam
Name: Francisco Campa


LAB 5 WRITE-UP

Procedure

Smart Phone Camera Settings

  • Type of Smartphone: IPhone 6
    • Flash: Off
    • ISO setting: 8
    • White Balance: Automatic
    • Exposure: Automatic
    • Saturation: Automatic
    • Contrast: Automatic


Calibration

  • Open phone's camera
  • Place upright in the cradle provided
  • Adjust the distance from phone to the sample making sure the picture is in focus and the distance is greater than 4 cm away.
Name: Set Up


  • Distance between the smart phone cradle and drop = 4.1 cm


Solutions Used for Calibration

Name: Water
Name: O.25
Name: 0.5
Name: 1
Name: 2
Name: 5



Placing Samples onto the Fluorimeter

  • 1. Pipet 80 milliliters of SYBR Green I solution in the middle of the 2 circles farthest away from the camera.
  • 2. Pipet 80 milliliters of calibration solution onto the first drop.
  • 3. Make sure the slide is lite by the blue light.


Data Analysis

Representative Images of Negative and Positive Samples

Name: Negative Sample
Name: Positive Sample

Image J Values for All Calibrator Samples


Concentration Area Mean RawIntDen Drop Raw IntDen Background Raw IntDen Drop- Background
0 582 30 17609 2428 15181
0.25 582 39.7 23098 6099 16999
0.5 582 39.6 23071 4984 18087
1 582 26.321 15319 909 14410
2 582 126.3 73506 4821 68685
5 582 112.4 65415 2478 62937


Calibration curve


PCR Results Summary

  • Our positive control PCR result was 5 μg/mL
  • Our negative control PCR result was 0 μg/mL

Observed results

  • Patient 73039 : The sample did not contain the allele so it did not glow green. The sample was observed to be 0 μg/mL.
  • Patient 40429 : The sample did contain the allele so it glowed green. The sample was observed to be 2 μg/mL.

Conclusions

  • Patient 73039 : This patient does not have coronary artery disease.
  • Patient 40429 : This patient does have coronary artery disease.



SNP Information & Primer Design

Background: About the Disease SNP

The disease SNP is associated with lipoprotein lipase, which helps regulate phospholipase activity. It caused by a genetic mutation in the 8th chromosome. More specifically, in the 5' strand, changing AAT to AGT. As a result, it causes coronary artery disease in homo sapiens. Coronary artery disease, or heart disease, is one of the deadliest diseases in America.

Primer Design and Testing

At the end of the primer test, the non-diseased primer test gave a 220bp reading, while the diseased primer had no match.


Name: Nondieased
Name: Diseased