BME103:T130 Group 6

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Contents

OUR TEAM

Name: Jocelynn ChristensenRole: Experimental Protocol Planner
Name: Jocelynn Christensen
Role: Experimental Protocol Planner
Name: Sam ZimmermanRole: OpenPCR Machine Engineer and Vampire Hunter Extraordinaire
Name: Sam Zimmerman
Role: OpenPCR Machine Engineer and Vampire Hunter Extraordinaire
Name: Adam HellandR&D
Name: Adam Helland
R&D
Name: Ryan UchimuraRole: Experimental Protocol Planner
Name: Ryan Uchimura
Role: Experimental Protocol Planner
Name: studentRole(s)
Name: student
Role(s)

LAB 1 WRITE-UP

Initial Machine Testing

The Original Design
Image:Open PCR Solid Works Design.png Image:Open PCR Solid Works Design 2.png


Experimenting With the Connections

When we unplugged the LED screen from the circuit board, the machine stopped displaying information on the LED screen.

When we unplugged the white wire that connects the circuit board to the heating block, the heating block would not heat up.


Test Run

(Write the date you first tested Open PCR and your experience(s) with the machine)




Protocols

Polymerase Chain Reaction
1. The Polymerase Chain Reaction or PCR works by singling out a single piece of DNA and then multiplying it so there's millions of copies of one strand of DNA. It's a step by step process that first occurs by heating up the DNA to 100°C in order to denature the hydrogen bonds between the two strands of DNA so that both sides of the DNA can be accesible to copy. After the strands are separated, specific primers are added to locate the section of DNA to be amplified. Next, the Taq DNA polymerase is added which actually copies the section of DNA desired and synthesizes the second half of each strand. After this there are only a few copies of the DNA which is why the machine then replicates more strands by repeating the process multiple times until there are millions of copies.

2.

3. The GoTaq master mix contains 400µM dATP, 400µM dGTP, 400µM dCTP, 400µM dTTP, and 3mM MgCl2



4.
Reagent Volume
Template DNA (20ng) 0.2μL
10μM forward primer 1.0μL
10μM reverse primer 1.0μL
GoTaq master mix 50.0μL
dH2O 47.8μL
Total Volume 100.0μL



5. Out of the eight samples we ran the PCR on, one was a positive control with the cancer DNA template, the other was a Negative control with no cancer DNA template. Then we had three samples of a 57 year old male's DNA (patient ID 19185). We also had three samples of a 63 year old female's DNA (patient ID 88142).


Flourimeter Measurements

(Add your work from Week 3, Part 2 here)




Research and Development

Specific Cancer Marker Detection - The Underlying Technology

  • Our genes can tell us anything and everything about ourselves.
    • The sooner we can detect cancer, the more effectively it can be prevented or treated.
So why not find out about our disposition to cancer with our genes?


The science community has identified many DNA sequences that are correlated to incidence of cancer. Through a process known as Polymerase Chain Reaction, (or PCR,) we can make tons of copies of any sequence of DNA from a DNA template. So, let's say we want to find out if someone has a sequence of DNA that may be indicative of a higher cancer risk; how can we do it?


r17879961 is a sequence of DNA that has been positively linked with cancer. It is a part of a sequence of DNA that codes for a protein kinase called CHEK2.



(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 OpenPCR tutorial might be useful. Be sure to credit the source if you borrow images.)




Results

(Your group will add the results of your Fluorimeter measurements from Week 4 here)


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