BME100 s2014:W Group12 L5: Difference between revisions
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'''Single-Drop Fluorimeter'''<br> | '''Single-Drop Fluorimeter'''<br> | ||
This lab uses a single drop fluoremeter device to detect DNA samples. This device consists of a sturdy outer cover which serves to block out interfering light sources so that the fluorescent light is more easily detected. We set up a phone cradle to secure the phone while it is taking pictures. Inside the device there is a platform for the slides to be placed on | This lab uses a single drop fluoremeter device to detect DNA samples. This device consists of a sturdy outer cover which serves to block out interfering light sources so that the fluorescent light is more easily detected. We set up a phone cradle to secure the phone while it is taking pictures. Inside the device there is a platform for the slides to be placed on a surface designed to repel water so droplets on the slide maintain a spherical shape. Around the slide there is a LED light source which emits a blue light in order to clearly see the fluorescing dye. | ||
'''How the Fluorescence Technique Works'''<br> | '''How the Fluorescence Technique Works'''<br> | ||
The blue LED light shines on the sample underneath the black box, in the dark, and detects the different DNA concentrations. | The blue LED light shines on the sample underneath the black box, in the dark, and detects the different DNA concentrations. The samples are shown to have different DNA concentrations based on the amount of light that they fluoresce. | ||
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'''Representative Images of Samples''' | '''Representative Images of Samples''' | ||
Image with No DNA[[Image:ImageJ with NO DNA.PNG]] | |||
Image with DNA[[Image:ImageJ with DNA.PNG]] | |||
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'''Fitting a Straight Line'''<br> | '''Fitting a Straight Line'''<br> | ||
[[Image:BME100_12_Calibration.jpg]] | |||
Latest revision as of 08:58, 16 April 2014
BME 100 Spring 2014 | 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-UPBackground InformationSYBR Green Dye
How the Fluorescence Technique Works
ProcedureSmart Phone Camera Settings
Calibration The phone is placed upright in the cradle with an eraser to help it stand straight. The cradle is moved about 6.5 cm away from the fluorimeter. This set up never moves once the experiment has begun.
Add more rows as needed
Step 1: Insert the slide, smooth side down Step 2: 80 milliliters of the SYBR GREEN on the first slide position Step 3: 80 milliliters of the calf thymus (water blank) on top of the "beach ball" of SYBR GREEN Step 4: Focus camera on the "beach ball", cover the fluorimeter with the black box, and take a picture in the dark Step 5: Repeat this process on different slide placements (at least three) for each sample.
Data AnalysisRepresentative Images of Samples
Image J Values for All Samples
Our positive control PCR result was 30.20 μg/mL. Our negative control PCR result was -36.08 μg/mL Patient 78086 : The images of the three samples of DNA given from Patient 78086 were without color. There was no sign of any SYBR Green 1 in all three images. The amount of corrected PCR product concentration was around -36 μg/mL. The exact values were: -35.89, -36.88 and -36.56 μg/mL. Patient 32445 : The images of the three samples of DNA given from Patient 32445 were without color as well. Again there was no sign of any SYBR Green 1 in all three images. The amount of corrected PCR product concentration was around -35 μg/mL. The exact values were: -35.23, -35.38 and -35.74 μg/mL. Patient 78086 : Negative. We were able to come to this conclusion based on the values of μg/mL for both the positive and negative controls. This patient's corrected PCR product concentration matched that of the negative control. Patient 32445 : Negative. We were able to come to this conclusion based on the values of μg/mL for both the positive and negative controls. This patient's corrected PCR product concentration matched that of the negative control as well. We did not get confused on the results and assume that the results of the patients would be different. They in fact were very similar and we were able to successfully ascertain the desired result.
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