BME100 s2015:Group2 12pmL5: Difference between revisions
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'''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 | * Our positive control PCR result was 0.0833 μg/mL | ||
* Our negative control PCR result was | * Our negative control PCR result was 0.0833 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 1 ID# 58575 : | ||
For all images of patient 1, the droplets all did not produce any type of green color when the drop samples were placed on top of the SYBR Green I dye. The drops themselves actually appeared to have a dark center. The concentration of DNA in these samples was the same as the positive and negative controls to which they were compared against. This is because the amount of PCR solution was 100μL which was then pipetted into the 500 μL solution given which resulted in 600 μL. After this solution was created, 80 μL was then pipetted onto the 80 μL of syber green dye which produced and amount which was 0.0833 μg/mL. Based upon the results of patient 1's data, they did not contain the diseased gene sequence. | |||
* Patient _____ : | * Patient _____ : | ||
Revision as of 16:28, 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
LAB 5 WRITE-UPProcedureSmart Phone Camera Settings
For the set-up, this picture represents what the set up looked like right before the group closed the flap to take the picture. The camera was set in the cradle and the lens was 6.8 cm away from the drop that was on the superhydrophic side of the glass slide. The box covered the Fluorimeter to protect the SyberGreen Dye from exposure to light. After close the flap, three pictures were taken using the timer on the iPhone to delay the taking of the pictures until after the flap was closed. Placing Samples onto the Fluorimeter
Data AnalysisRepresentative Images of Negative and Positive Samples Negative Sample Positive Sample
Image J Values for All Calibrator Samples In the calibration section of the chart listed above, picture 1 correlates to H2O, picture 2 correlates to 0.25 units of DNA per microliter, picture 3 correlates to 0.5 units of DNA per microliter, picture 4 correlates to 1.0 units of DNA per microliter, picture 5 correlates to 2.0 units of DNA per microliter, and picture 6 correlates to 5.0 units of DNA per microliter. Picture 6 served as the positive control while picture 1 served as the negative control when checking the results of the PCR experiment.
Observed results
For all images of patient 1, the droplets all did not produce any type of green color when the drop samples were placed on top of the SYBR Green I dye. The drops themselves actually appeared to have a dark center. The concentration of DNA in these samples was the same as the positive and negative controls to which they were compared against. This is because the amount of PCR solution was 100μL which was then pipetted into the 500 μL solution given which resulted in 600 μL. After this solution was created, 80 μL was then pipetted onto the 80 μL of syber green dye which produced and amount which was 0.0833 μg/mL. Based upon the results of patient 1's data, they did not contain the diseased gene sequence.
Conclusions
SNP Information & Primer DesignBackground: About the Disease SNP
Primer Design and Testing The diseased primer had no matches because the humane genome does not have the mutation of AGT. Therefore, there would be no match in accordance with the natural human genome tested in the website. While polymorphisms, unique alleles and variations are all what helps to make each individual have a slightly unique genome, the AGT mutation is what can lead to a person having coronary heart disease. The diseased primer has no results while the healthy, non-diseased primer produces a result. Diseased Primer <a href="http://imgur.com/ruzkEYS"><img src="http://i.imgur.com/ruzkEYS.png" title="source: imgur.com" /></a> NON-Diseased Primer <a href="http://imgur.com/TAV9kPT"><img src="http://i.imgur.com/TAV9kPT.png" title="source: imgur.com" /></a>
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