BME103:W930 Group5 l2

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Contents

OUR TEAM

Name: Andrea CarpenterRole(s): Experimental Protocol Planner
Name: Andrea Carpenter
Role(s): Experimental Protocol Planner
Name: Malik McLaurinRole(s): Open PCR Machine Engineer
Name: Malik McLaurin
Role(s): Open PCR Machine Engineer
Name: Dana McElwainRole(s): Open PCR Machine Engineer
Name: Dana McElwain
Role(s): Open PCR Machine Engineer
Name: Chris AnastosRoles(s): R&D Scientist
Name: Chris Anastos
Roles(s): R&D Scientist
Name: StudentRole(s)
Name: Student
Role(s)

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
The following picture is a simple change in the design of the large exterior screw located on the top of the Open PCR lid:
[IMG:screw.png]


Instructions





Protocols

Materials

Supplied in Kit

Supplies Amount
Micro Test Tubes?
Transfer Pipettes?
PCR Master Mix6 samples (598.8μL total)
Positive Control Solution*1 sample (100.0μL***)
Negative Control Solution**1 sample (100.0μL***)
PCR Machine1
Fluorimeter1

(*)Positive control consists of calf thymus DNA
(**)Negative control simply consists of a blank solution of water
(***)Already mixed with PCR master mix

Included in Fluorimeter Package:

Supplies Amount
Smart phone stand1
LCD Box1
Light box1
Sybr green solution500.0μL

Components of PCR master mix:

DNA Solution Component Amount
Patient’s Template DNA*0.2μL
10μM forward primer1.0μL
10μM reverse primer1.0μL
Promega GoTaq master mix50.0μL
dH2O47.8μL
Total100.0μL

(*)Not actually included in kit, but must be added to the master mix by the user.

Supplied by User

Supplies Amount
Smart Phone with Camera1
Patient's Template DNA6 samples (0.2μL each)
External Computer1
Image J Software1
Open PCR Software1
Gloves1 pair
Lab Coat1


PCR Protocol



DNA Measurement Protocol

Research and Development

Background on Disease Markers

For this experiment, our group chose to take an in-depth look at acute myeloid leukemia (AML). AML is a type of cancer that begins inside the bone marrow. The immune system of the human body is ultimately affected by AML, as bone marrow helps fight infections. The white blood cells that grow and form in bone marrow are turned into cancerous cells; the cells grow very quickly and sporadically, thus replacing healthy white blood cells. Our reference single nucleotide polymorphism associated with acute myeloid leukemia is rs121912500. In this SNP, the pathogenic allele for AML is classified as a single nucleotide variation. This means that only one nucleotide is altered in the allele causing AML. This variation results in a missense mutation.




Primer Design



Illustration


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