BME100 s2014:T Group13 L5: Difference between revisions
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'''SYBR Green Dye'''<br> | '''SYBR Green Dye'''<br> | ||
SYBR Green dye was added to each of the samples from the PCR on the sample slides. | SYBR Green dye was added to each of the samples from the PCR on the sample slides. A drop of each sample was placed onto a drop of this green dye each time the Fluorimeter was used. These drops of dyes react with different molecules of DNA. In the samples which had no DNA, these reactions didn’t take place. The reaction between the DNA and the green dye cause the blue excitation light to be absorbed and a green light to be emitted. Thus the samples with DNA shone green and the samples without DNA simply shone the original blue from the excitation light. The exterior black box, or light box, is used in order keep all other light from interfering with the images being captured. | ||
'''Single-Drop Fluorimeter'''<br> | '''Single-Drop Fluorimeter'''<br> | ||
The Fluorimeter allows for the quantitative evaluation of samples that underwent a Polymerase Chain Reaction (PCR). It is made up of an interior black box, or table, plastic trays, a slide port, a blue excitation light, a camera phone cradle, a Samsung galaxy s4 camera phone, and an exterior black box that covers all other components. The first component, the interior black box, basically acts as both a table for the slide port and a house for the blue excitation light. The plastic trays sit under this interior box and allows researchers to correctly line up the slide samples and the camera they are using. The slide port is the narrow canal in which the sample slides sit. This port allows for the all samples to be easily interchanged | The Fluorimeter allows for the quantitative evaluation of samples that underwent a Polymerase Chain Reaction (PCR). It is made up of an interior black box, or table, plastic trays, a slide port, a blue excitation light, a camera phone cradle, a Samsung galaxy s4 camera phone, and an exterior black box that covers all other components. | ||
The first component, the interior black box, basically acts as both a table for the slide port and a house for the blue excitation light. The plastic trays sit under this interior box and allows researchers to correctly line up the slide samples and the camera they are using. The slide port is the narrow canal in which the sample slides sit. This port allows for the all samples to be easily interchanged.The blue excitation light is a light that is shone through the samples. The next component is the camera phone cradle and the camera phone (Samsung Galaxy s4). The camera phone is used to take high quality pictures of the samples so that they can then be transferred to a computer. The cradle simply holds the camera in one place. The last component is the exterior black box. This box covers all other components and keeps all unwanted light away from the experiment. | |||
[[Image:Fluorimter.PNG|390px]] | [[Image:Fluorimter.PNG|390px]] |
Revision as of 11:12, 17 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
The first component, the interior black box, basically acts as both a table for the slide port and a house for the blue excitation light. The plastic trays sit under this interior box and allows researchers to correctly line up the slide samples and the camera they are using. The slide port is the narrow canal in which the sample slides sit. This port allows for the all samples to be easily interchanged.The blue excitation light is a light that is shone through the samples. The next component is the camera phone cradle and the camera phone (Samsung Galaxy s4). The camera phone is used to take high quality pictures of the samples so that they can then be transferred to a computer. The cradle simply holds the camera in one place. The last component is the exterior black box. This box covers all other components and keeps all unwanted light away from the experiment.
ProcedureSmart Phone Camera Settings
Calibration
[Instructions: See worksheet page 6.]
[Add more rows as needed]
Data AnalysisRepresentative Images of Samples
[Instructions: See worksheet page 8. To save time on typing a new Wiki table from scratch, use THIS TOOL to auto-generate a Wiki table: Excel-to-Wiki Converter. Copy the headers and values from the Excel spreadsheet you made, paste them into the form field, click submit, copy the Wiki code that the tool generated, and replace TABLE GOES HERE (below) with your auto-generated code.]
Instructor's summary: You completed 8 PCR reactions in a previous lab. You 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 concnetration of claf thymus DNA) to convert INTDEN values into DNA concentration. Your positive control and negative control samples are used to determine the threshold values for determining whether an unknown (patient) sample is truly positive or negative.
Compare each patient's results to the positive control value and the negative control value. Draw a final conclusion for each patient (positive or negative) and explain why you made that conclusion.
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