BME103:W930 Group2: Difference between revisions
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=LAB 1 WRITE-UP= | =LAB 1 WRITE-UP= | ||
<br>DISCLAIMER: ALL GROUP MEMBERS PARTICIPATED EQUALLY IN THE WRITING OF REPORTS. SOME OF US HAD ISSUES LOGGING IN SO CONTRIBUTIONS MAY NOT APPEAR TO ADD UP APPROPRIATELY. | |||
==Initial Machine Testing== | ==Initial Machine Testing== | ||
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Step 1: Denaturation by Heat <br> | Step 1: Denaturation by Heat <br> | ||
Initially, heat separates a DNA strand into two separate strands. This is allowed because the hydrogen bonds | Initially, heat separates a DNA strand into two separate strands. This is allowed because the hydrogen bonds that hold the DNA together are weak and easily separable when heated. <br> | ||
Step 2: Annealing Primer to Target Sequence<br> | Step 2: Annealing Primer to Target Sequence<br> | ||
We want to target a sequence specifically and in order to accomplish that you must use primers. Primers mark the end of the target sequence. Two primers are included in the PCR; one for each of the strands that were just separated during denaturation. The beginning of the DNA target sequence is also marked by the primers that bind (anneal) to the complementary sequence. <br> | We want to target a sequence specifically and in order to accomplish that you must use primers. Primers mark the end of the target sequence. Two primers are included in the PCR; one for each of the strands that were just separated during denaturation. The beginning of the DNA target sequence is also marked by the primers that bind (anneal) to the complementary sequence. <br> | ||
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1. A PCR Machine is created to amplify a patient’s DNA sample. In order to use a PCR machine, one must first gather the DNA samples, 50 μL each. <br> | 1. A PCR Machine is created to amplify a patient’s DNA sample. In order to use a PCR machine, one must first gather the DNA samples, 50 μL each. <br> | ||
2. The frozen DNA samples are | 2. The frozen DNA samples are placed into separate tubes and once melted, 8 transfer pipettes are used to add primer. <br> | ||
3. Once the samples were ready, we processed the DNA in the openPCR system. <br> | 3. Once the samples were ready, we processed the DNA in the openPCR system. <br> | ||
Latest revision as of 17:05, 10 December 2012
BME 103 Fall 2012 | Home People Lab Write-Up 1 Lab Write-Up 2 Lab Write-Up 3 Course Logistics For Instructors Photos Wiki Editing Help | ||||||||||||||||||||||||||||||||||||||||||
OUR TEAMLAB 1 WRITE-UP
Initial Machine Testing
When we unplugged part mounting plate from circuit board, Machine #2 had no visible screen and we could not see the the information on the LCD screen. When we unplugged the white wire that connects the circuit board to the sample holder, the machine's temperature was unable to be monitored and could not be regulated.
Our group did our test run on October 24, 2012. During the test run, we ran into no problems with Machine #2. We were done and found our results in a matter of 1 to 2 hours. The Open PCR machine was connected to one computer to control the temperature of each cycle of PCR during the experimental.
ProtocolsPolymerase Chain Reaction How PCR Works: Step 1: Denaturation by Heat
1. A PCR Machine is created to amplify a patient’s DNA sample. In order to use a PCR machine, one must first gather the DNA samples, 50 μL each.
400μM dATP, 400μM dGTP, 400μM dCTP, 400μM dTTP and 3mM MgCl2, reaction buffer (pH 8.5), nuclease-free water
Fluorimeter assembly procedure: Turn on the excitation light on the fluorimeter. Put a smart phone in the cradle and set it up to take pictures of the slide. Place two drops of water in the middle of the first two rows of the slide using a pipette. Align the drop by moving the slide so the drop is in the middle of the black fiber optic fitting. Cover the fluorimeter with the light box while maintaining the ability to use the smart phone to take pictures.
How to open images in Image J: Save the pictures to the phone. Download the pictures onto a computer that has Image J. Open them with Image J by going to add image. Research and DevelopmentSpecific Cancer Marker Detection - The Underlying Technology Cancer Associated with the gene: Breast and Colorectal cancer with a susceptibility to LiFraumeni Syndrome Source: http://omim.org/entry/604373?search=CHEK2&highlight=chek2
Cancer Gene Sequence: Baye's Rule:
Results
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