BME100 s2015:Group7 9amL4

BME 100 Spring 2015	Home People 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 Photos Wiki Editing Help
OUR TEAM Name: Luke Shaffer Role(s) Name: Brandon Hansen Role(s) Name: Harris Tran Role(s) Name: Jaid Coronel Role(s) LAB 4 WRITE-UP Protocol Materials Lab Coat Disposable Gloves PCR reaction mix 8 tubes, 50 μL each contains Taq DNA polymerase, MgCl2, and dNTP’s DNA/ primer mix, 8 tubes, 50 μL each A strip of empty PCR tubes Disposable pipette tips Cup for discarded tips Micropipettor OpenPCR machine PCR Reaction Sample List Tube Label PCR Reaction Sample Patient ID G7 + Positive control none G7 - Negative control none G7 1-1 Patient 1, replicate 1 71504 G7 1-2 Patient 1, replicate 2 71504 G7 1-3 Patient 1, replicate 3 71504 G7 2-1 Patient 2, replicate 1 96389 G7 2-2 Patient 2, replicate 2 96389 G7 2-3 Patient 2, replicate 3 96389 DNA Sample Set-up Procedure Check that group has all required materials Cut strip of empty PCR tubes in half, resulting in two strips of four linked tubes Label the sides of the empty tube with tube labels, making sure to not mark the lids Place tubes in rack Transfer 50 μL of PCR reaction mix into the tube labeled positive control, disposing of the tip into the collection cup afterwards With a new pipette tip, transfer the positive control DNA/primer mix into the same tube. Repeat steps 5 and 6 for the negative control, patient 1 replicates 1,2, and 3, and patient 2 replicates 1,2, and 3. Seal the lids of the PCR reaction tubes tightly. Take the tubes to the PCR machine, and place them into the heating block. Return all reusable materials to the appropriate storage places. Dispose of all biomedical material waste in an orange/ red Biohazard Bag. OpenPCR program The thermal cycling program is a program that is used to control the OpenPCR machine. Using a varied set of input data, the program can tell the machine to heat up to a specific temperature, cool down to a specific temperature, and specify the number of times to repeat the cycle. Research and Development PCR - The Underlying Technology The purpose of the PCR (Polymerase Chain Reaction) is to replicate a small strand of DNA sample into a batch large enough to measure through technological means. The DNA sample taken from a patient acts as the template for the replication to base from; meaning after the process of the chain reaction, multiple amounts of the patient's DNA is measurable. While primers in the chemical solution acts as the beginning placeholders for Taq polymerase to add complementary nucleic acids (such as Adenine, Thymine, Cytosine, Guanine) to the DNA template. The deoxyribonucleotides (dNTP's) acts as the building materials in this solution, for the polymerase to attach onto the separated DNA strand. These are the main components for PCR to perform effectively and efficiently.