Overview of the Original Diagnosis System Through the use of the polymerase chain reaction, it is possible to test the DNA of patients in order to determine if the disease associated SNP is in their DNA. The patient DNA was isolated and placed into a solution with a large amount of the base pairs, TAQ polymerase, and specific primers that correspond to the coding area of the disease associated SNP. All thirty-four lab groups were responsible for two patients, which means that 68 strands of DNA were tested. Many processes were implemented to ensure a minimal amount of error. In each lab group, each patient’s DNA was tested three times; this means that there were a total of 204 polymerase chain reactions run. In order to ensure that the PCR machine was working correctly, controls were included with the samples of DNA from the patients. These controls were known to be positive and negative, respectively, for the disease-associated SNP. Once these PCR was completed the results were analyzed with a fluorescent agent that was activated in the presence of the disease-associated SNP. The level of fluorescence was analyzed in an image analysis program called ImageJ, using images taken from an IPhone. For every trial three images were taken to ensure no error. This means that, with all of the lab groups taken into consideration, 612 images were analyzed in ImageJ. These are only images from the patients and don’t include the analysis of the controls and known concentrations. All of this data was then collected and compiled into a large spreadsheet for the entire class of BME 100 students. The data was fairly accurate most likely due to all of the efforts made to ensure that error was minimal.
What Bayes Statistics Imply about This Diagnostic Approach
Computer-Aided Design
TinkerCAD
TinkerCAD is an online product design service that allowed us to replicate the structure of the OpenPCR machine. Using the building blocks given to us through the website thingiverse.com, we re-created a PCR pachine virtually. We then modified this basic structure to incorporate some of the changes we felt would better the device.
Our Design
Feature 1: Consumables Kit
For our new device, the consumable products will come premeasured to ensure accuracy. A set of eight PCR test tubes containing the predetermined 50 μL of PRC mixture (containing the TAQ polymerase) will be provided along with pre-labeled test tubes with 50 μL of DNA. Besides this, the kit will come with the necessary pipette, sterile pipette tips, and gloves.
The inclusion of premeasured consumables is meant to prevent any human error in the PCR process. With the tubes of 50 μL PCR mix, the individual will only have to pipette the DNA samples into the mix and not worry about the amount of PCR mixture. Besides this, the test tubes, both for the primer mixture and DNA are labeled to prevent any confusion or cross contamination. The tubes for both the PCR mixture and DNA will correspond.
Feature 2: Hardware - PCR Machine & Fluorimeter
Increase the speed of pcr machine and have flourimeter do imagej analysis
BME 100 Fall 2014
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