BME100 s2015:Group4 9amL5

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BME 100 Spring 2015 Home
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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: Measho Habtemichael
Name: student
Name: student
Name: student
Name: Amanda Nguyen


LAB 5 WRITE-UP

Procedure

Smart Phone Camera Settings

  • Type of Smartphone: Samsung Galaxy S5
    • Flash: OFF
    • ISO setting: 800
    • White Balance: AUTO
    • Exposure: +2.0
    • Saturation: N/A
    • Contrast: N/A

Calibration

  • Distance between the smart phone cradle and drop = 5 cm


Solutions Used for Calibration

initial concentration of 2X calf thymus DNA solution (micrograms/mL) volume of the 2X DNA solution (microliters) Volume of the SYBR GREEN I Dye solution (microliters) Final DNA concentration in SYBR Green I solution (micrograms/mL)
5 80 80 2.5
2 80 80 1
1 80 80 0.5
0.5 80 80 0.125
0.25 80 80 0.125
0 80 80 0



Placing Samples onto the Fluorimeter

  1. In the center of the first two rows of the slide, put 80 microliters of SYBR GREEN I
  2. Ensure that the drop is centered
  3. Add 80 micrometers of water
  4. Move the slide so that the drop is aligned and focused by the blue LED to the middle of the black fitting opposite the drop
  5. Cover the drop with the light box
  6. Use the timer take a picture of the drop
  7. Take three images of the focused drop
  8. Remove the box
  9. Remove the sample using the pipettor
  10. Repeat steps for next 5 samples


Data Analysis

Representative Images of Negative and Positive Samples


Image J Values for All Calibrator Samples


TABLE GOES HERE


Calibration curve


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 _____ :




SNP Information & Primer Design

Background: About the Disease SNP

SNP is a DNA sequence that is associated with coronary heart and artery disease.

Primer Design and Testing

In the first part of designing the primers, it was necessary to validate the two non disease primers that were designed using the instructions from the lab workbook. The primers tested were as follows.
Non disease froward primer: AATCTGGGCTATGAGATCAA
Non disease reverse primer: GAAACACCAGGGCTCAGGGT

This is the information inputted into the UCSC In-Silico PCR website

These are the results of the input.

Then in the second part of designing, disease forward and reverse primers were created. The primers tested were as follows:
Disease forward primer: AATCTGGGCTATGAGATCAG
Disease reverse primer: TTAGACCCGATACTACTAGTT

This is the information inputted into the website

These are the results of the input

From the results, it can be seen that there are no matches to these primers. This is because they are being tested using a non-disease human genome sequence. The disease primers are being compared against a genome that do not have disease.