BME100 f2013:W1200 Group16 L4

BME 100 Fall 2013	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: John Jakoubek Name: Courtney Willson Name: Mychal Hooser Name: Ayanna Akinyemi Name: Rene Reynolds LAB 1 WRITE-UP Initial Machine Testing The Original Design A Polymerase Chain Reaction (PCR) Machine takes a DNA sequence and makes millions of copies of that sequence. This is done by the experimenter placing the DNA that they desire to copy into the test tube along with DNA polymerase, 2 primers, magnesium chloride, and nucleotides. Once placed in the test tubes and into the machine, the machine has to go through 3 heated steps. The first step is to heat the test tubes to 95 degrees Celsius. This denatures the 2 DNA strands. It is then cooled to 57 degrees Celsius which allows the primers to attach to the appropriate DNA sequences. It is then heated to 72 degrees Celsius. This allows the DNA polymerase to attach to the primers and begin replication. These three steps are repeated 35 times resulting in multiple copies of the desired DNA. Experimenting With the Connections When we unplugged the LCD cable from the OpenPCR LCD, the display screen turned off so the readings were no longer available. When we unplugged the lid temperature sensory wire from the OpenPCR LCD, the machine's temperature of the heating block changed from 26.1 C to -40.0 C. Test Run The first test run was on October 23, 2013 at 12:58 p.m. At 1:31 p.m. the machine was still showing the temperature at 30.8 degrees Celsius even though it began at 26 degrees Celsius. It was also only on cycle 1. Therefore, it is concluded that this machine is not functioning properly. Wednesday, October 23, 2013 Protocols Thermal Cycler Program DNA Sample Set-up PC 1 2 3 NC A B C Patient 1: 41731 Patient 2: 78042 DNA Sample Set-up Procedure Place the PCR reaction mix into each individual tube using a pipette Add in DNA primer to the tubes using a clean pipette tip, being sure not to cross-contaminate the samples. Place tubes into the PCR machine and close the lid. PCR Reaction Mix What is in the PCR reaction mix? The PCR reaction mix is made up of Taq DNA polymerase, MgCl2 (magnesium chloride), and dNTP's (nucleotides). The polymerase functions to replicate the DNA strand. The magnesium chloride functions as a cofactor as the process cannot proceed without its presence. The dNTP's are the prime nucleotides as they function as building blocks to produce the synthesized DNA strands. DNA/ primer mix What is in the DNA/ primer mix? The DNA/primer mix is made up of different templates of DNA. The mix consists of the same forward primer and reverse primer. Research and Development PCR - The Underlying Technology (Add a write-up, essay-style, organized into paragrpahs with descriptive headers, based on the Q&A's from Section three of your worksheet) (BONUS points: Use a program like Powerpoint, Word, Illustrator, Microsoft Paint, etc. to illustrate how primers bind to the cancer DNA template, and how Taq polymerases amplify the DNA. Screen-captures from the PCR video/ tutorial might be useful. Be sure to credit the sources if you borrow images.)