BME100 f2014:Group6 L5

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Contents

OUR TEAM

Name: Prakriti Shukla
Name: Prakriti Shukla
Name: Gavin White
Name: Gavin White
Name: Galt Goettl
Name: Galt Goettl
Name: Laura Stokes
Name: Laura Stokes
Name: Paul Chua
Name: Paul Chua
Name: Yamilex Bustamante
Name: Yamilex Bustamante


LAB 5 WRITE-UP

Procedure

Smart Phone Camera Settings

  • Type of Smartphone: iPhone 5
    • Flash: Off
    • ISO setting:n/a
    • White Balance: n/a
    • Exposure:n/a
    • Saturation:n/a
    • Contrast:n/a


Calibration

  • Distance between the smart phone cradle and drop = 4 cm


Solutions Used for Calibration

Concentration ofCalf Thymus DNA solution(micrograms/mL) Volume of theDNA solution (μL) Volume of the SYBR GREEN I Dye Solution (μL) Final DNA concentration in SYBR Green I solution (μg/mL)
5 80 80 2.5
2 80 80 1
1 80 80 0.5
0.5 80 80 0.25
0.25 80 80 0.125
0 80 80 0



Placing Samples onto the Fluorimeter

  1. "[Place the slide, rough side up, onto the fluorimeter]"
  2. "[Place 80 microliters of the SYBR Green Solution into the two middle circles of the slide]"
  3. "[Then place 80 microliters of the calibration solution into the same drop of the SYBR Green solution]"
  4. "[Adjust the slide so the drop is lighted by the blue light of the fluorimeter]"


Data Analysis

Representative Images of Negative and Positive Samples

Positive Sample Image:Positivecontrolgroup6.png Negative Sample Image:Negativecontrolgroup6.png


Image J Values for All Calibrator Samples

Image:PCRPRODUCT1.png Image:PCRPRODUCT2.png Image:PCRPRODUCT3.png Image:PCRPRODUCT4.png

Calibration curve
Image:Concentrationgraphgroup6.png

PCR Results Summary

  • Our positive control PCR result was 5.44 μg/mL
  • Our negative control PCR result was -3.19 μg/mL

Observed results

  • Patient 29148: The first sample was very green
  • Patient 35264 : The second sample had no color

Conclusions

  • Patient 29148: This patient was close to the positive control and would amplify nearly as much as the positive control.
  • Patient 35264 : This patient was close to the negative control and will not amplify as much




SNP Information & Primer Design

Background: About the Disease SNP

SNP is a single nucleotide polymorphism. It occurs commonly within a population in which a single nucleotide (A, T, C, G) differs between members of a biological species. Almost all common SNPS occur within two alleles. SNPs occur within non-coding regions rather than coding regions. Genomic recombination and mutation rate can also vary SNP density. These genetic variations are exploited in DNA fingerprinting which is used in forensic science

Primer Design and Testing The non-disease primer was successful while the disease primer did not have any results because of the SNPs in the disease primer. Image:Nondiseaseprimerfinal.png Image:Diseaseprimerfinal.png

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