BME103:T130 Group 5 l2: Difference between revisions
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# Invert tubes, then turn upright to mix solution. | # Invert tubes, then turn upright to mix solution. | ||
# Plug in OpenPCR Machine and turn it on. | # Plug in OpenPCR Machine and turn it on. | ||
# Open lid and place tubes into holder in PCR machine. | # Open lid and place tubes into holder in PCR machine (the machine can now hold a maximum of 24 tubes). | ||
# Close the lid. | # Close the lid. | ||
# Add the following cycles on the OpenPCR program: | # Add the following cycles on the OpenPCR program: |
Revision as of 21:48, 28 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.
Key Features
Instructions
ProtocolsMaterials
PCR Protocol
Photo origin: http://openwetware.org/wiki/BME103:T130_Group_9
Research and DevelopmentBayesian statistics
A=Alzheimer's B=Positive test Result [math]\displaystyle{ P(A/B)=P(B/A)P(A)/P(B) }[/math]
Background on Disease Markers Alzheimer's disease is a form of dementia that occurs with loss of brain function. It affects multiple areas of the brain associated with memory, language, personality, perception, and cognitive skills. The disease typically manifests itself through forgetfulness, but gradually progresses to inability to perform basic functions, speak, and recognize family members. Currently, there is no cure. Treatment tries to slow down the disease or at the least, manage symptoms. An SNP related to Alzheimer's disease is rs1466662 (http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=1466662). It is located on chromosome four, the intron region of NM_001142552.1 and arises from a missense mutation replacing an A with a T. It is the most significant SNP outside of the SNP linked to APOE.
The backwards primer is TAT TTT TAG AAG CGA TAA AA. The forwards primer is GCC TCT TTG CCC TCT GTT TT. An allele not containing the disease will not have the sequence that allows the primers to bind. If the primers cannot bind, then that means Taq polymerase does not know where to bind. If Taq polymerase does not bind, then the sequence does not get replicated. Therefore, there will be no PCR product. Conversely, if the disease allele is present, the primers will bind. Taq polymerase will then be able to bind to the DNA and replicate the strands, creating more double-stranded DNA yielding a PCR product.
The wanted gene in the figure above refers to rs1466662. Photo courtesy of http://users.ugent.be/~avierstr/principles/pcr.html
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