BME100 s2015:Group17 12pmL5: Difference between revisions

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'''Background: About the Disease SNP'''
'''Background: About the Disease SNP'''
<!-- 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. -->
 
SNP, single nucleotide polymorphism is a DNA sequence variation occurring within a small population where a single base, A, T, C, or G differs between paired chromosomes or biological species. SNP are one of the most important genetic mutations that impact common disease. SNP results from replication errors and DNA damage, this phenomenon occurs exactly once in human evolution.  Only sometimes does SNP have a correlation to a certain disease or trait. SNP has several applications in medicine such as gene discovery, allele mapping, and drug response prediction. 


'''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. -->
 
Results obtained from this lab were used to analyze the DNA sequence of two patients, one with a disease and the other without a disease. This lab demonstrated that primers bind to a certain region of a DNA sequence in order for the amplification of a small sample of DNA.  Amplification occurs in two different directions on the DNA strand, the 5` and 3` ends. The 5` primers begins its sequence at the origin of the disease SNP variation location of 34370656. This was the location in the human genome where a thymine nucleotide has been mutated to a cytosine nucleotide, such a mutation results in the SNP disease that is seen in one of the patients. We used the UCSC In-Silico PCR website to test the non-disease primer to match the rs19956218 sequence.
 
[[Image:Ss+(2015-04-01+at+02.47.54).png]]





Revision as of 15:20, 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: Hannah Austin
Name: Warner Kostes
Name: Ivanna Revel
Name: Alexandria Morales
Name: Your name


LAB 5 WRITE-UP

Procedure

Smart Phone Camera Settings

  • Type of Smartphone: iPhone 6
    • Flash: Inactive
    • ISO setting: N/A (setting not able to be adjusted)
    • White Balance: N/A (setting not able to be adjusted)
    • Exposure: N/A (setting not able to be adjusted)
    • Saturation: N/A (setting not able to be adjusted)
    • Contrast: N/A (setting not able to be adjusted)


Calibration

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


Solutions Used for Calibration

Initial Concentration of 2X Calf Thymus DNA solution (micrograms/mL) Volume of the 2X DNA solution (μL) Volume of the SYBR GREEN I Dye solution (μL) Final DNA concentration in SYBR Green I soluton (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. [Instructions: Step one, in your OWN words]
  2. [Instructions: Step two, in your own words]
  3. [Instructions: Step three, in your own words]
  4. [Instructions: Step etc., in your own words]


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, single nucleotide polymorphism is a DNA sequence variation occurring within a small population where a single base, A, T, C, or G differs between paired chromosomes or biological species. SNP are one of the most important genetic mutations that impact common disease. SNP results from replication errors and DNA damage, this phenomenon occurs exactly once in human evolution. Only sometimes does SNP have a correlation to a certain disease or trait. SNP has several applications in medicine such as gene discovery, allele mapping, and drug response prediction.

Primer Design and Testing Results obtained from this lab were used to analyze the DNA sequence of two patients, one with a disease and the other without a disease. This lab demonstrated that primers bind to a certain region of a DNA sequence in order for the amplification of a small sample of DNA. Amplification occurs in two different directions on the DNA strand, the 5` and 3` ends. The 5` primers begins its sequence at the origin of the disease SNP variation location of 34370656. This was the location in the human genome where a thymine nucleotide has been mutated to a cytosine nucleotide, such a mutation results in the SNP disease that is seen in one of the patients. We used the UCSC In-Silico PCR website to test the non-disease primer to match the rs19956218 sequence.