BME100 f2013:W1200 Group1 L5: Difference between revisions

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{| {{table}} width=700
{| {{table}} width=700
|-
|-
| row 1 cell 1 || row 1 cell 2 || row 1 cell 3 || row 1 cell 4
| Calf Thymus DNA solution concentration (microg/mL) || Volume of the 2x DNA solution (µL)|| Volume of the SYBR Green I Dye solution (µL) || Final DNA concentration in SYBR Green I Assay (ng/mL)
|-
|-
| row 2 cell 1 || row 2 cell 2 || row 2 cell 3 || row 2 cell 4
| 5 || 80 || 80 || 2.5
|-
|-
| row 3 cell 1 || row 3 cell 2 || row 3 cell 3 || row 3 cell 4
| 2 || 80 || 80 || 1
|-
|-
| row 4 cell 1 || row 4 cell 2 || row 4 cell 3 || row 4 cell 4
| 1 || 80 || 80 || 0.5
|-
|-
| row 5 cell 1 || row 5 cell 2 || row 5 cell 3 || row 5 cell 4
| 0.5 || 80 || 80 || 0.25
|-
|-
| row 6 cell 1 || row 6 cell 2 || row 6 cell 3 || row 6 cell 4
| 0.25 || 80 || 80 || 0.125
|-
|-
| row 7 cell 1 || row 7 cell 2 || row 7 cell 3 || row 7 cell 4
| 0 || 80 || 80 || blank
|}
|}
''[Add more rows as needed]''
''[Add more rows as needed]''

Revision as of 14:10, 6 November 2013

BME 100 Fall 2013 Home
People
Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
Course Logistics For Instructors
Photos
Wiki Editing Help

OUR TEAM

Name: Chloe Kwon
Name: Jeff Miles
Name: Casey Weinstein
Name: Nima Afzalian
Name: Mario Gonzalez


LAB 5 WRITE-UP

Background Information

SYBR Green Dye
SYBR Green Dye is a small molecular dye that does not fluoresce well in water or with single strands of DNA, but fluoresces very well with dsDNA. It is combined with the DNA solution in order to detect fluorescence.


Single-Drop Fluorimeter
The single-drop fluorimeter detects fluorescence within the drop of DNA solution. It measures the wavelength distribution and intensity when light passes through the DNA solution. Add a PHOTO for bonus points


How the Fluorescence Technique Works
The fluorescence technique works by combining a fluorescent SYBR green dye with the DNA solution. When the blue light beam from the single-drop fluorimeter, the drop will turn a fluorescent green color indicating the presence of DNA. The droplet's color will be detected by the smartphone and the picture will then be analyzed using the ImageJ software.



Procedure

Smart Phone Camera Settings

  • Type of Smartphone: HTC One
    • Flash: off
    • ISO setting: 800
    • White Balance: Auto
    • Exposure: Neurtral
    • Saturation: Neutral
    • Contrast: Lowest Setting


Calibration

[Instructions: Describe how to set up your camera in front of the fluorimeter. Add a PHOTO of this set-up for bonus points.]

  • Distance between the smart phone cradle and drop =

[Instructions: See worksheet page 6.]


Solutions Used for Calibration [Instructions: See worksheet page 6.]

Calf Thymus DNA solution concentration (microg/mL) Volume of the 2x DNA solution (µL) Volume of the SYBR Green I Dye solution (µL) Final DNA concentration in SYBR Green I Assay (ng/mL)
5 80 80 2.5
2 80 80 1
1 80 80 0.5
0.5 80 80 0.25
0.25 80 80 0.125
0 80 80 blank

[Add more rows as needed]


Placing Samples onto the Fluorimeter

  1. [Instructions: Step one, in your OWN words]
  2. [Instructions: Step two, in your own words]
  3. [Instructions: Step three, in your own words]
  4. [Instructions: Step etc., in your own words]


Data Analysis

Representative Images of Samples

[Instructions: Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with no DNA]

[Instructions: Show an IMAGE where you drew a circle around the droplet with the freehand tool for a sample with DNA (positive signal)]


Image J Values for All Samples

[Instructions: See worksheet page 8. To save time on typing a new Wiki table from scratch, use THIS TOOL to auto-generate a Wiki table: Excel-to-Wiki Converter. Copy the headers and values from the Excel spreadsheet you made, paste them into the form field, click submit, copy the Wiki code that the tool generated, and replace TABLE GOES HERE (below) with your auto-generated code.]


TABLE GOES HERE


Fitting a Straight Line

[Instructions: Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 9]