BME103:T130 Group 17 l2

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|dH<sub>2</sub>O || 95.6 μL
|dH<sub>2</sub>O || 95.6 μL
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|'''Total Volume'''|| 199.6 μL (+ DNA = 200 μL)
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|'''Total Volume'''|| 199.6 μL  
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{| {{table}}
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|'''Supplied by the user''' || '''Amount'''
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|DNA || (40 ng) 0.4 μL
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|Computer || 1
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|Lab coats || 1 per person
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|gloves || 2 per person
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|}
'''PCR Protocol'''
'''PCR Protocol'''

Revision as of 15:47, 28 November 2012

BME 103 Fall 2012 Home
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Lab Write-Up 1
Lab Write-Up 2
Lab Write-Up 3
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Contents

OUR TEAM

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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
Image of the heating block

Image of the cooling system


Key Features
The first picture shows the heating block that is in use in the current PCR machine. It is limited in that it can only contain 16 test tubes at any given time. Our redesign calls for doubling the amount of test tube holders by increasing the size of the heating block. The picture below is that of the current cooling system. Because our redesign is going the be running PCR on twice as many samples, an increase in the cooling power needs to occur in order to sufficiently cool the larger number of samples. This is done by increasing the size of the cooling system and therefore increasing its power.


Instructions
1) Remove the current heating block and cooling system from the PCR machine.
2) Replace removed items with the larger heating block and larger cooling system.
3) Replace side panels with larger ones in case any of the replaced parts exceed the confines of the current PCR machine.
4) Continue to run PCR as normal.




Protocols

Materials

Materials Table

Supplied in the Kit Amount
Pre-labeled PCR tubes 32 tubes
Pre-labeled bulb pipets 34 pipets
20 µM forward primer (ALZ) 2.0 μL
20 µM reverse primer (ALZ) 2.0 μL
GoTaq Master Mix 100.0 μL
dH2O 95.6 μL
Total Volume 199.6 μL
Supplied by the user Amount
DNA (40 ng) 0.4 μL
Computer 1
Lab coats 1 per person
gloves 2 per person

PCR Protocol



DNA Measurement Protocol 1. Turn on the Blue led light. 2. Place the smart phone in the cradle provided at the right angle and distance from the slide. 3. Turn on the camera on the phone. TURN OFF THE FLASH. Set the ISO to 800 or higher. Increase the exposure to maximum. 4. Adjust the distance between phone and the first two rows of dots of the slide so that the picture will be clear and the camera does not give a blurred image. 5.First label the blank pipettes according to the patients (A,B,C,D... all eight of them). The pipettes given by the instructor are color coded. The white coded pipette is used for water. the red coded pipette is used for the calibrator (the tube with the red dot). The blue coded pipette is used for the sybrgreen (the tube with the blue dot). The black coded pipette is used to pick up the waste and put it in the cup that collects the waste droplets.

Research and Development

Background on Disease Markers



Primer Design



Illustration


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