BME103:W930 Group5 l2: Difference between revisions

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Line 69: Line 69:
| Transfer Pipettes||?
| Transfer Pipettes||?
|-
|-
| DNA Solution||6 samples (600.0μL)
| PCR Master Mix||6 samples (598.8μL total)
|-
|-
| Positive Control Solution||1 sample (100.0μL)
| Positive Control Solution*||1 sample (100.0μL***)
|-
|-
| Negative Control Solution||1 sample (100.0μL)
| Negative Control Solution**||1 sample (100.0μL***)
|-
|-
| PCR Machine||1
| PCR Machine||1
Line 79: Line 79:
| Fluorimeter||1
| Fluorimeter||1
|}<br>
|}<br>
*Positive control consists of calf thymus DNA<br>
**Negative control simply consists of a blank solution of water<br>
***Already mixed with PCR master mix


<i>Included in Fluorimeter Package</i><br>
<i>Included in Fluorimeter Package</i><br>
Line 105: Line 109:
| 10μM reverse primer||1.0μL
| 10μM reverse primer||1.0μL
|-
|-
| GoTaq master mix||50.0μL
| Promega GoTaq master mix||50.0μL
|-
|-
| dH<sub>2</sub>O||47.8μL
| dH<sub>2</sub>O||47.8μL
|-
|-
| Total||100.0μL
| Total||100.0μL
|}<br>
*Not actually included in kit, but must be added to the master mix by the user.
<b>Supplied by User</b><br>
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Supplies'''
| align="center" style="background:#f0f0f0;”|'''Amount'''
|-
| Smart Phone with Camera||1
|-
| DNA Solution||6 samples (0.2μL each)
|-
| External Computer||1
|-
| Image J Software||1
|-
| Open PCR Software||1
|-
| Gloves||1 pair
|-
| Lab Coat||1
|}<br>
|}<br>


Line 115: Line 141:




'''PCR Protocol'''
<u>'''PCR Protocol'''</u>








'''DNA Measurement Protocol'''
<u>'''DNA Measurement Protocol'''</u>


==Research and Development==
==Research and Development==

Revision as of 23:19, 25 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

OUR TEAM

Name: Student
Role(s): Experimental Protocol Planner
Name: Student
Role(s)
Name: Student
Role(s)
Name: Student
Role(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


Instructions





Protocols

Materials

Supplied in Kit

Supplies Amount
Micro Test Tubes ?
Transfer Pipettes ?
PCR Master Mix 6 samples (598.8μL total)
Positive Control Solution* 1 sample (100.0μL***)
Negative Control Solution** 1 sample (100.0μL***)
PCR Machine 1
Fluorimeter 1

  • 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 stand 1
LCD Box 1
Light box 1
Sybr green solution 500.0μL

Components of PCR master mix:

DNA Solution Component Amount
Patient’s Template DNA* 0.2μL
10μM forward primer 1.0μL
10μM reverse primer 1.0μL
Promega GoTaq master mix 50.0μL
dH2O 47.8μL
Total 100.0μL

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

Supplied by User

Supplies Amount
Smart Phone with Camera 1
DNA Solution 6 samples (0.2μL each)
External Computer 1
Image J Software 1
Open PCR Software 1
Gloves 1 pair
Lab Coat 1


PCR Protocol



DNA Measurement Protocol

Research and Development

Background on Disease Markers



Primer Design



Illustration