BME103:T130 Group 10 l2

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Research and Development)
(Protocols)
Line 145: Line 145:
'''PCR Protocol'''  
'''PCR Protocol'''  
<br>
<br>
 +
1.) Be sure to be in an area with natural lighting for use of the solar battery, or locate an area with a standard power source availible <br>
 +
2.) Connect the PCR machine via USB to the computer <br>
 +
4.)
'''DNA Measurement Protocol'''
'''DNA Measurement Protocol'''

Revision as of 15:00, 29 November 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
Image:BME494_Asu_logo.png

Contents

OUR TEAM

Name: Frances LakersRole:Protocol Planner
Name:
Frances Lakers
Role:
Protocol Planner
Name: Tyler TamasauckasRole:Protocol Planner
Name:
Tyler Tamasauckas
Role:
Protocol Planner
Name: Jeffery RamirezRole:R&D Specialist
Name:
Jeffery Ramirez
Role:
R&D Specialist
Name: Alexander Baldwin Role:Machine Design
Name:
Alexander Baldwin
Role:
Machine Design

LAB 2 WRITE-UP

Thermal Cycler Engineering

Our re-design is based upon the Open PCR system originally designed by Josh Perfetto and Tito Jankowski.


System Design


Key Features

There are two specific features that will be upgraded in the OpenPCR machine - the case and the power source. The case will be changed from a thin wood to a more durable plastic, so that the device can be taken out into the field. The device will also have solar panels, which will provide for the power. This shift allows the device to be used where electricity is not always available, such as rural and undeveloped regions. When combined, these two features open the device up to a larger market.

Instructions

The instructions would remain the same, as the solar panel array would come attached to the plastic on the device's top side.




Protocols

Materials

Supplied in Kit

Item Quantity
PCR Machine 1 unit
Camera Phone Stand 1 unit
Fluorimeter Stand (LED Light) 1 unit
BlackOut Box 1 unit
Operator's Manual 1 unit

Also included: Access code for smart phone app download.

Supplied by User

Item Quantity
Smart Phone (w/ Camera) 1 unit
USB/Phone Cord (if desired) 1 unit
Micropipettes (as needed)
Hydrophobic Glass Slides (as needed)
Eppendorf Tubes Max 16 tubes/trial
Sample DNA 0.2μL/tube; 16 tubes/trial
GoTAQ Master Mix 50μL/tube; 16 tubes/trial
10 μM Forward Primer 1μL/tube; 16 tubes/trial
10 μM Reverse Primer 1μL/tube; 16 tubes/trial
Distilled H2O 47.8μL/tube; 16 tubes/trial
SYBR Green 0.1mL/test sample

PCR Protocol
1.) Be sure to be in an area with natural lighting for use of the solar battery, or locate an area with a standard power source availible
2.) Connect the PCR machine via USB to the computer
4.) DNA Measurement Protocol

Research and Development

Background on Disease Markers

The disease that we chose was Sickle Cell Anemia. Sickle Cell is a inherited disease that comes from both parents. If only one parent has the Sickle Cell trait, then the child will be a carrier. If both parents have the trait, then the child will have Sickle Cell. This disease causes the red blood cells to change into a crescent shape instead of being disk shape resulting in the loss of efficient oxygen delivery. It can also result in clotting because the blood cells can not make it through small blood vessels very well. The SNP for this disease is 78478128 and the reference SNP is rs78478128.

The link to the page with more info on Sickle cell is http://www.ncbi.nlm.nih.gov/pubmedhealth/PMH0001554/

The link to the page with the genetic information and SNP is http://www.ncbi.nlm.nih.gov/projects/SNP/snp_ref.cgi?rs=78478128#locus


Primer Design

The forward primer for Sickle Cell is 5'-tcctaagccagtgccagaag-3'

The reverse primer for Sickle Cell is 5'-gaattcgtctgtttcccattctaaac-3'

The mutation looks like this: TCCTGAGGAG with the bold a being the mutation.

A disease allele will give a PCR result and a non-disease allele will not because PCR is designed to amplify the area that corresponds to the mutation on the DNA. If it did not amplify only the target region, there would be too much information and a result would not be possible.



Illustration


Image:PCR_process.gif


Personal tools