BME103 s2013:T900 Group4 L2: Difference between revisions

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'''Calibration''' Photo of Fluorimeter  
'''Calibration'''  
 
Photo of Fluorimeter  


* Put your phone on the cradle at a right angle
* Put your phone on the cradle at a right angle

Revision as of 18:03, 1 April 2013

BME 103 Spring 2013 Home
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Lab Write-Up 1
Lab Write-Up 2
Lab Write-Up 3
Course Logistics For Instructors
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OUR TEAM

Name: Kinjal Ahir : Procedure
Name: Tuan Phan : Background Information
Name: student
Name: student
Name: student


LAB 2 WRITE-UP

Background Information

SYBR Green Dye
[A short summary describing SYBR green dye]

SYBR Green I is a fluorescent dye that is effectively used to bind double stranded DNA.

Single-Drop Fluorimeter
[A description of the single-drop fluorimeter device. Add a PHOTO for bonus points]

Fluorimeter is a device that is used to measure fluorescence.

How the Fluorescence Technique Works
[In your own words, a summary of the information from page 9 of the worksheet]



Procedure

Smart Phone Camera Settings

  • Iphone 4
    • Flash: Off
    • ISO setting:NA
    • White Balance: NA
    • Exposure: NA
    • Saturation: NA
    • Contrast: NA



Calibration

Photo of Fluorimeter

  • Put your phone on the cradle at a right angle
  • Then fix the height of the fluorimeter so that the camera can take picture of the drop on sideways

  • Distance between the smart phone cradle and drop = 4cm


Solutions Used for Calibration

Calf Thymus DNA solution concentration (microg/ML) Volume of the 2x DNA solution (uL) Volume of the SYBR GREEN Dye Solution (uL) Final DNA concentration PicoGreen 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


Placing Samples onto the Fluorimeter

  • Step one, Check your phone camera setting, and turn the flash off
  • Step two,Pick up DAN, gloves, lab coat, glass slide, buffer tubes, sync green I tube, and DNA standard
  • Step three, Slide the glass slide into fluorimeter
  • Step four, Drop 80 micro liters of sample of the solution where the light is flashing
  • Step five, Put the box around it and take pictures of the drops

Data Analysis

Representative Images of Samples

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

[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 [See worksheet page 5]

Trial DNA Concentration (μg/mL) Area Mean Pixel Value RAWINTDEN (drop) RAWINTDEN (background)
1 0 10,906 21.672 236,351 70,338
1 0.25 10,044 35.971 361,290 42,572
1 0.5 10,562 84.274 890,103 52,974
1 0.5 10,452 50.546 528,303 50,012
1 1 10,350 115.226 1,192,594 39,995
1 2 10,914 193.145 2,107,989 46,923
1 5 10,282 142.563 1,465,554 55,342
2 0 10,808 50.412 544,856 71,106
2 0.25 10,510 51.038 536,412 39,795
2 0.5 10,688 78.799 842,203 57,075
2 1 10,432 90.965 948,944 52,390
2 2 10,103 152.075 1,536,481 61,000
2 5 10,440 115.674 1,207,635 51,873
3 0 10,131 37.264 377,520 66,289
3 0.25 10,448 61.987 647,645 48,887
3 0.5 10,762 64.475 693,875 40,978
3 1 10,465 74.835 783,144 47,549
3 2 10,188 155.63 1,585,554 43,186
3 5 10,672 72.312 771,711 47,142

[Add more rows as needed]


Fitting a Straight Line
[Place an IMAGE of your Excel plot with a line of best fit here. See worksheet page 8]