BME100 f2013:W1200 Group2 L4: Difference between revisions
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<u>Procedure:</u><br> | <u>Procedure:</u><br> | ||
Step 1:Determine positive and negative control. The negative control will not have the cancer marker. The positive control has cancer marker. | Step 1: Determine positive and negative control. The negative control will not have the cancer marker. The positive control has cancer marker.<br> | ||
Step 2:Label Tubes. <br> | Step 2: Label Tubes. <br> | ||
Positive control: cancer negative control: Not cancer<br> | Positive control: cancer negative control: Not cancer<br> | ||
Patient 1 43849 Patient 2 80853 <br> | Patient 1 43849 Patient 2 80853 <br> | ||
1A 2A<br> | 1A 2A<br> | ||
1B | 1B 2B<br> | ||
1C 2C<br> | 1C 2C<br> | ||
Step 3:Add DNA/primer mix to positive control.<br> | Step 3: Add DNA/primer mix to positive control.<br> | ||
Step 4:Add PCR reaction mix to positive control.<br> | Step 4: Add PCR reaction mix to positive control.<br> | ||
Step 5:Place sample in Open PCR(thermo cycler)<br> | Step 5: Place sample in Open PCR(thermo cycler)<br> | ||
Step 6:While in the Open PCR the sample will be heated up to 95℃ the DNA chain separates into two strands. The temperature then decreases to 50℃ and the primers attach to the separate strands at the target sequence. The polymerase attaches to the primer and begins to replicate the strand.<br> | Step 6: While in the Open PCR the sample will be heated up to 95℃ the DNA chain separates into two strands. The temperature then decreases to 50℃ and the primers attach to the separate strands at the target sequence. The polymerase attaches to the primer and begins to replicate the strand.<br> | ||
Step 7:Repeat steps 3-5 for the remaining tubes.<br> | Step 7: Repeat steps 3-5 for the remaining tubes.<br> | ||
Step 8:Compare Patient 1 DNA replicates to those of the positive and negative controls.<br> | Step 8: Compare Patient 1 DNA replicates to those of the positive and negative controls.<br> | ||
step 9:Compare Patient 2 DNA replicates to those of the positive and negative controls. | step 9: Compare Patient 2 DNA replicates to those of the positive and negative controls. | ||
Revision as of 11:46, 26 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-UPThe Original Design Experimenting With the Connections When we unplugged (part 3) from (part 6), the machine screen went blank. (did what? fill in your answer) When we unplugged the white wire that connects (part 6) to (part 2), the machine read -40 degrees Celsius on the screen. (did what? fill in your answer)
(Write the date you first tested Open PCR and your experience(s) with the machine)
ProtocolsThermal Cycler Program
DNA Sample Set-up Procedure Materials: Procedure: Positive control: cancer negative control: Not cancer Step 3: Add DNA/primer mix to positive control.
PCR Reaction Mix
-Taq DNA Polmerase
Research and DevelopmentPCR - The Underlying Technology PCR Reaction Components The components of the PCR Reaction include the template DNA, primers, Taq Polymerase, Magnesium Chloride(MgCl2), and Deoxyribonucleotides (dNTP's). Each component or reagent is added to the PCR tube before running them through the PCR device and software called "Open PCR". The template DNA is the source of genetic information in which an enzyme copies from. The Taq Polymerase is an enzyme and specific kind of DNA Polymerase that is thermostable. It reads the genetic information form the DNA template and checks for mutations before copying data. Thermal Cycling Thermal cycling is the process of cycling a PCR tube holding PCR reaction in specific variations of time and temperature. The PCR reaction is first exposed to high temperatures, it is lowered, and increases slightly in temperature. The changes in temperature triggers and activates specific enzymes and/or reagents and is done ... Initial Step Denature Anneal Extend Final Step Final Hold Nucleotides and Base-pairing (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.)
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