BME100 s2015:Group5 12pmL4

OUR TEAM

Name: Nick Valverde

Name: Daniella Orlando

Name: Brandon Favre

Name: Martisha Clyde

Name: Fernanda Diva

Name: David Shumate

Lab A

Section 1: Protocol
Materials

• Lab coat and disposable gloves
• PCR reaction mix, 8 tubes, 50 μL containing Taq DNA polymerase, MgCl2, and dNTP's
• DNA/primer mix, 50 μl each: all have the same forward and reverse primer but different templates of DNA
• A strip of empty PCR tubes
• Disposable pipette tips: only use each only once. Never re-­‐use disposable pipette tips or samples will be cross-­‐contaminated
• Cup for discarded tips
• Micropipettor
• OpenPCR machine

DNA Sample Set-up Procedure

1. Obtain patient samples, including a positive and negative control, from the TA as well as the patient ID's for group
2. Record which ID's went with each patient and label each sample with the ID consistent with the patient
3. Do not write the ID's on the tubes, only record on the table
4. Add the polymerase, nDTP, and primers to each of the 8 samples
5. The samples are prepared for PCR

Section 2: Research and Development-the Underlying Technology Each component of PCR serves a functional component:

• Template DNA is a guide to extend to the primer, which produces new double stranded DNA.
• Primers copy DNA sequences by attaching to either end of the strand, and allow polymerases to start their DNA replication duties.
• Taq Polymeraseis the enzyme responsible for utilizing the surrounding nucleotides to synthesize new strands of DNA based on the template DNA strand
• Deoxyribonucleotides (dNTP's)are nucleotides that are bonded to deoxyribose, which is itself bonded to a phosphate. These provide the structural building blocks for DNA replication.