BME100 f2013:W1200 Group15 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 |100px|thumb|Name: Patrick McFarland Protocol Planning]] Name: Zac Roy Open PCR Machine Testing Name: Taylor Olvey Open PCR Machine Testing Name: Gregory Berghorst Research and Development Name: Tameem Jamal Research and Development LAB 1 WRITE-UP Initial Machine Testing The Original Design Source(https://www.google.com/search?q=Open+PCR+machine&source=lnms&tbm=isch&sa=X&ei=ZyxoUsWsO6bwiwLEooCYAw&ved=0CAcQ_AUoAQ&biw=1920&bih=961#facrc=_&imgdii=_&imgrc=Ct82Mj8rloOURM%3A%3BYGkEtXwApqO_fM%3Bhttp%253A%252F%252Fopenwetware.org%252Fimages%252F4%252F41%252FPCR_Machine.png%3Bhttp%253A%252F%252Fopenwetware.org%252Fwiki%252FBME103%253AT130_Group_7%3B1377%3B1027). (Add image of the full OpenPCR machine here, from the Week 9 exercise.) This device is known as the OpenPCR is used for the amplification of DNA. The samples that are placed in the machine consist of DNA, two different primers, DNA polymerase, and nucleotides. Minimal DNA is needed for the reaction so hair and small blood samples would suffice. Once a proper PCR sample is ready, it is put inside to machine where it will be put through a cycle of heating and cooling. The sample will then be heated to 95 degrees Celsius, causing the DNA to "unzip". It is then cooled down to 50 degrees Celsius so that the primers can react with the DNA. Heated up once again to 72 degrees Celsius, the Dna polymerase and begins to add complimentary nucleotides to the stands. This cycle will repeat 30 times until there are over a billion copies of the desired DNA. Experimenting With the Connections When we unplugged (part 3) from (part 6), the machine ... Will not be able to monitor the heat source that is being applied to the DNA samples. (did what? fill in your answer) When we unplugged the white wire that connects (part 6) to (part 2), the machine ... Lossed all power and would not heat up the DNA samples to the corect tempature. (did what? fill in your answer) Test Run The date we first tested Open PCR was October 23, 2013. The machine is fairly simple and easy to work with. The computer program for Open PCR was fast pace and very understandable. Little if no confusion usi ng this machine occured. The Open PCR machine creates a new experiment fairly quickly, and has absolutly no difficulty to operate the device. (Write the date you first tested Open PCR and your experience(s) with the machine) Started at 12:32pm and by 1:32pm was at cycle 17. Protocols Thermal Cycler Program DNA Sample Set-up Positive Control: PCC Patient 1 Sample 1: P11 Patient 1 Sample 2: P12 Patient 1 Sample 3: P13 Negative Control: NCC Patient 2 Sample 1: P21 Patient 2 Sample 2: P22 Patient 2 Sample 3: P23 DNA Sample Set-up Procedure 1. Receive 8 sample tubes from Professor, containing 50μL of PCR reaction material 2. Label Samples according to table, to avoid swapping results 3. Put appropriate DNA sample into the accordingly labeled tube. Use a new pipette tip for each test tube to avoid cross contamination 4. Place the 8 sample tubes into the Thermocycler 5. Follow Thermocycler instructions above PCR Reaction Mix 1. Taq DNA polymerase 2. MgCl2 3. dNTP's DNA/ primer mix 1. Sample of patient's DNA 2. Forward Primer 3. 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.)