Brent Hayes Russon: Research and Development Specialist
Abdulaziz Alamal: Open PCR Machine Engineer
Andrew Munoz: Experimental Protocol Planner
Abdullah Alqahtani: Open PCR Machine Engineer
LAB 2 WRITE-UP
Thermal Cycler Engineering
Our re-design is based upon the Open PCR system originally designed by Josh Perfetto and Tito Jankowski.
System Design
Key Features
Instructions
Protocols
Materials
PCR Protocol
DNA Measurement Protocol
Research and Development
Background on Disease Markers
The marker that is being used is rs137852453. This SNP is associated with hemophiia. Hemophilia is a condition where someone's blood is not clotting properly. Data on this particular SNP variance can be seen here: http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=137852453
The associated gene change goes from a CGG (healthy gene sequence) to TGGG (sequence associated with hemophilia).
The gene alteration leads to a mutated human protein. It goes from R[Arg] to W[Trp]
Primer Design
Bottom Primer (in reverse):
TGGAATTGG T GGGTGGAAT
Top Primer (forward):
TTCAAAGCTTCAAGTATATC
These primers will attach to the other half of the DNA, but only if there is a matching genetic code for them to attach to. The top primer should always have a matching pair to attach to because there should be no genetic variance in its counterpart while the bottom primer will only match up with its corresponding strand of DNA if the genetic variance is present. A Taq DNA polymerase then connects to any attached primers, which along with MgCl2, makes it possible for free-floating nucleotides to fill in the rest of the letters missing from the DNA strand. This process is then repeated numerous times. If the mutation is not present then only the top primer will find a match and the reproduction of DNA will not show a noticeable increase. If the mutation is present in the subject's DNA then both primers will find matching pairs and create two full new sets of this sequence. As the process is repeated the amount of the sequences present will increase exponentially.
BME 103 Fall 2012
