BME100 f2013:W1200 Group18 L4: Difference between revisions
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| [[Image:Srtgsdhgf.jpg|100px|thumb|Name: Carlyn Harris<br>Role(s):Protocol]] | | [[Image:Srtgsdhgf.jpg|100px|thumb|Name: Carlyn Harris<br>Role(s):Protocol]] | ||
| [[Image:BME103student.jpg|100px|thumb|Name: Tori Platt<br>Role(s):Reseach and Development]] | | [[Image:BME103student.jpg|100px|thumb|Name: Tori Platt<br>Role(s):Reseach and Development]] | ||
| [[Image: | | [[Image:Nikhil.jpg|100px|thumb|Name: Nikhil Patel<br>Role(s):Initial PCR Testing]] | ||
| [[Image:BME103student.jpg|100px|thumb|Name: Alexandra Olson<br>Role(s): Protocol]] | | [[Image:BME103student.jpg|100px|thumb|Name: Alexandra Olson<br>Role(s): Protocol]] | ||
| [[Image:BME103student.jpg|100px|thumb|Name: Matthew Armas<br>Role(s):Initial PCR Testing]] | | [[Image:BME103student.jpg|100px|thumb|Name: Matthew Armas<br>Role(s):Initial PCR Testing]] |
Revision as of 19:14, 29 October 2013
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 TEAMLAB 1 WRITE-UPInitial Machine TestingTis is a PCR machine. The sample holder is at the top of the machine, this is where we will place tubes with samples in them to be held in place by holes in the square. The heating lid covers the sample holder, it is there to keep the heat around the sample holder. The heater inside the PCR machine will heat up the sample holder. The fan is there to keep the inside of the PCR machine cool, because there are electronics that can be damaged from over heating. The circuit board keeps the sensors active in a screen on top of the PCR machine, it tellls us how hot the heater will be, how long the machine will run for the experiment, and other data as well.
When we unplugged (part 3) from (part 6), the machine's screen on top of the box no longer functioned. The screen shows the data of the experiment, but because it was unplugged, we could not see the data. When we unplugged the white wire that connects (part 6) to (part 2), the machine's heat sensor no longer functioned. It determines how hot the machine runs, therefore the machine did not pick up any heat when it was unplugged. Test Run Open PCR was first tested on 10/23/13 from 1:18pm to 1:44pm. During the lab, we inspected the 16-tube PCR block and the heating lid, which are both designed to retain a constant temperature across the samples that would be tested. We also explored the insides of the machine, and how its various components affected one another. Our experience with the machine was somewhat frustrating because the machine failed to complete the required amount of cycles in a reasonable amount of time. It was labeled as being slow.
ProtocolsThermal Cycler Program
DNA Sample Set-up
PCR Reaction Mix The PCR reaction mix contains: -Tag DNA Polymerase -MgCl2 -dNTP's
DNA/ primer mix The DNA primer mix contains: -Different template DNAs -Forward primer -Reverse primer
Research and DevelopmentPolymerase Chain Reaction (PCR) - The Underlying Technology Polymerase chain reaction occurs when DNA is heated and cooled at exact temperatures for exact intervals of time. However, there are five components required for the reaction to work successfully; template DNA, primers, TAQ polymerase, Magnesium Chloride (MgCl2), and dNTPs (deoxyribonucletides). In order to replicate a section of DNA, a source for the sequence needs to be present for the primers to attach to. The template DNA provides the original copy to begin the entire process. What PCR enables is the replication of a target sequence in DNA that is made thousands of times over. Once a target DNA sequence is established, primers are responsible for marking the beginning and end of these sequences in order for them to be coded in the right direction (5'-3') by Taq Polymerase. Primers are short strands of oglionucleotides that tell the enzyme where to stop and start. After primers have attached themselves to the template DNA, an enzyme called Taq Polymerase binds t Deoxyribonucleotides (dNTPs) are essentially the building blocks of DNA. Each nucleotide base pairs with one and only one other base. For example, guanine only pairs with cytosine while thymine only pairs with adenine in double stranded DNA. After Taq Polymerase latches itself onto the location of the primers on template DNA or a previously created strand, it beings to attach dNTPs to compliment whichever base pair comes up as it goes along the sequence. In the Polymerase Chain Reactions, deoxyribonucleotides are present for new strands of DNA to be created throughout the process. (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.)
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