BME103:T130 Group 11 l2: Difference between revisions
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First, DNA samples were obtained from 31 different patients, each giving 3 DNA samples each. These DNA samples were stored in a freezer in microtubes. For patient number 1, his or her DNA samples were labelled 1a, 1b and 1c, using the heatproof marker. The other DNA samples from the remaining 30 patient were labelled in the same way. After all the DNA samples were obtained, each sample was transferred over to PCR tubes, which are tubes that are used especially for the PCR process. Each PCR tube was labelled in the same way as before. In addition to these 93 PCR tubes, two more tubes, each containing a negative control and a positive control respectively. The PCR tube containing the negative control was labelled "-" and the PCR tube containing the positive control was labelled "+". Using separate pipettes for each DNA sample and the controls, Primer 1 was added to every PCR tube. Once Primer 1 is added, Primer 2 is added to all the tubes as well, with separate pipettes still being used for all transfers. AFter these two primers are added, nucleotides are added in the same way. Finally, DNA Polymerase is added to all the PCR tubes. | First, DNA samples were obtained from 31 different patients, each giving 3 DNA samples each. These DNA samples were stored in a freezer in microtubes. For patient number 1, his or her DNA samples were labelled 1a, 1b and 1c, using the heatproof marker. The other DNA samples from the remaining 30 patient were labelled in the same way. After all the DNA samples were obtained, each sample was transferred over to PCR tubes, which are tubes that are used especially for the PCR process. Each PCR tube was labelled in the same way as before. In addition to these 93 PCR tubes, two more tubes, each containing a negative control and a positive control respectively. The PCR tube containing the negative control was labelled "-" and the PCR tube containing the positive control was labelled "+". Using separate pipettes for each DNA sample and the controls, Primer 1 was added to every PCR tube. Once Primer 1 is added, Primer 2 is added to all the tubes as well, with separate pipettes still being used for all transfers. AFter these two primers are added, nucleotides are added in the same way. Finally, DNA Polymerase is added to all the PCR tubes. | ||
All 95 of the PCR tubes are placed into the PCR machine, and the software for the PCR machine was opened on the computer that the PCR machine was connected to. | All 95 of the PCR tubes are placed into the PCR machine, and the software for the PCR machine was opened on the computer that the PCR machine was connected to. | ||
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'''DNA Measurement Protocol''' | '''DNA Measurement Protocol''' | ||
The fluorimeter was set up according to the image shown below. | |||
[[Image:BME103_Group11_FluorimeterProtocol.JPG|200px| A complete setup of the Fluorimeter ]] | |||
==Research and Development== | ==Research and Development== |
Revision as of 19:56, 27 November 2012
BME 103 Fall 2012 | Home People Lab Write-Up 1 Lab Write-Up 2 Lab Write-Up 3 Course Logistics For Instructors Photos Wiki Editing Help | ||||||||||||||||||||||||||||||||||||||||
OUR TEAMLAB 2 WRITE-UPThermal Cycler EngineeringOur re-design is based upon the Open PCR system originally designed by Josh Perfetto and Tito Jankowski.
In order to increase tube space, we increase the heating plate and increase the power proportionally to the size of the plate. We would also add more ventilation next to the heating plate in order to speed up the cooling process of the DNA. Key Features
ProtocolsMaterials
PCR Protocol
Research and DevelopmentBackground on Disease Markers More information on the gene can be found at http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=35731153
Illustration
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