BME100 s2014:T Group13 L5: Difference between revisions
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'''Placing Samples onto the Fluorimeter''' | '''Placing Samples onto the Fluorimeter''' | ||
The first step to introducing the samples onto the Fluorimeter was having the Cyber Green solution, the varying samples of DNA samples, two micropipettes each set at 80 micro liters, a plethora of pipette tips, and having the slide positioned in the correct place. | |||
The second step was to put 80 micro liters of the Cyber Green in the center of the slide so that the Blue Excitation Light would shine through it, using one micropipette and discarding the tip afterwards into a cup. | |||
The third step was putting 80 micro liters of a determined sample onto the 80 micro liters of Cyber Green solution with the other micropipette, discarding the tip afterwards into the same cup. | |||
The fourth step was to take a photo using the Samsung Galaxy S4, while having the exterior black box fully closed. | |||
The fifth step was to use one of the micropipettes to take off the combined Cyber Green, DNA solution from the slide, discarding the solution into a specified cup. | |||
The sixth and final step was to move the slide so that contamination wouldn’t happen. | |||
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Revision as of 11:14, 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]
The second step was to put 80 micro liters of the Cyber Green in the center of the slide so that the Blue Excitation Light would shine through it, using one micropipette and discarding the tip afterwards into a cup. The third step was putting 80 micro liters of a determined sample onto the 80 micro liters of Cyber Green solution with the other micropipette, discarding the tip afterwards into the same cup. The fourth step was to take a photo using the Samsung Galaxy S4, while having the exterior black box fully closed. The fifth step was to use one of the micropipettes to take off the combined Cyber Green, DNA solution from the slide, discarding the solution into a specified cup. The sixth and final step was to move the slide so that contamination wouldn’t happen.
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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