BME100 s2016:Group3 W1030AM L4
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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
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LAB 4 WRITE-UP
PCR Reaction Sample List
FINAL STEP: 72°C for 2 minutes
Research and Development
PCR - The Underlying Technology
"Template DNA:" the non-coding strand of a gene in DNA "Primers:" a strand of short nucleic acid sequences that serve as the starting point for DNA replication "Taq Polymerase:" a method for greatly amplifying short segments of DNA "Deoxyribonucleotides (dNTP’s):" the monomer, or single unit, of DNA, or deoxyribonucleic acid. (contains components for PCR)
2.What happens to the components (listed above) during each step of thermal cycling?
“INITIAL STEP”: Add the DNA, primers, polymerase, and nucleotides to one test tube. “Denature”: The double stranded DNA template breaks apart when heated and becomes a single strand, because the weak hydrogen bonds break in higher temperatures. “Anneal”: At this lower temperature, the DNA strands want to pair together again. However, the primer sequences crowd the DNA and attach. “Extend”: When the temperature is increased to 72°C, the polymerase are activated and attach to the primer and DNA. The polymerase act as organizers for the nucleotides and add them to the single strands of DNA to their complementary nucleotide (A-T and G-C). “FINAL STEP”: Repeat these steps 20-30 times by raising and lowering the temperature to allow the DNA to replicate. During the third cycle, your desired segments of DNA start to appear. “FINAL HOLD”: Slows the DNA replication process to a near stop in order for it to be examined as a preserved image.
3. Adenine always binds to Thymine forming one pair, and Guanine and Cytosine form the other base pair.
SNP Information & Primer Design
Background: About the Disease SNP
"Non-Hodgkin's Lymphoma Summary:"
Non-Hodgkin's Lymphoma or NHL, is a series of blood cancers that include every type of blood cancer except Hodgkin's Lymphoma. It can be passed down genetically, or it can be caused by exposure to certain toxic chemicals, autoimmune disease, HIV and Hepatitis, and through over exposure to radiation. The five year survival rate for this form of cancer is 69% and every 9 minutes, one person dies from a form of Non-Hodgkin's Lymphoma, which is 155 each day. Treatments for this type of cancer are chemotherapy, radiation therapy, STEM-cell transplants, and specialized medications. This cancer shows up on the body in the form of tumors on the lymph nodes. Abdominal and chest pain are also common symptoms of Non-Hodgkin's Lymphoma.
Primer Design and Testing