BME103 s2013:T900 Group9 L2

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(Background Information)
(Background Information)
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==Background Information==
==Background Information==
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[[Image:sdp.png|400px|]]<br>
'''SYBR Green Dye'''<br>
'''SYBR Green Dye'''<br>

Revision as of 20:23, 1 April 2013

BME 103 Spring 2013 Home
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Lab Write-Up 1
Lab Write-Up 2
Lab Write-Up 3
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Contents

OUR TEAM

Name: Coley White
Name: Coley White
Name: Aimen Vanood
Name: Aimen Vanood
Name: Brady Falk
Name: Brady Falk
Name: student
Name: student


LAB 2 WRITE-UP

Background Information


SYBR Green Dye
SYBR Green Dye is a type of dye that is used to highlight when dsDNA is present because that is when it fluoresces the strongest. The dye fluoresces weak with water or single strands of DNA. [A short summary describing SYBR green dye]


Single-Drop Fluorimeter
The Single-Drop Fluorimeter is a custom device designed for the purpose of finding when a double-stranded piece of DNA is present. It fluoresces double-stranded DNA fragments in a green color, and does nothing to a solution without the double-stranded piece of DNA. [A description of the single-drop fluorimeter device. Add a PHOTO for bonus points]


How the Fluorescence Technique Works
The process had several different variables that were chosen for very specific reasons. The slide to be used was chosen because it had a rough surface with glass circles exposed in it, causing the solution to concentrate at those locations and be easier to examine. The DNA dye chosen for the experiment was used because it was not completely soluble, which allowed for the light to glow instead of causing a reflective light off of the drop. [In your own words, a summary of the information from page 9 of the worksheet]



Procedure

Smart Phone Camera Settings

  • Type of smart phone used and adjusted camera settings.
    • Phone: HTC Droid Incredible 2
    • Flash: None
    • ISO setting: 800
    • White Balance: Auto
    • Exposure: +2
    • Saturation: +2
    • Contrast:-2


Calibration

[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 = 3.7 cm

[See worksheet page 5]


Solutions Used for Calibration [See worksheet page 5] Image J Values for All Samples [See worksheet page 5]

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

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, in your OWN words]
  • [Step two, in your own words]
  • [Step three, in your own words]
  • [etc.]


Data Analysis

Representative Images of Samples

Sample with no DNA
Description of image

Sample with DNA (positive signal)
Description of image



Calf Thymus DNA

concentration (FINAL),

migrog/mL
Area Mean Pixel Value

RAWINTDEN

of the drop

RAWINTDEN

of the background

Water 40636 32.437 1318101 302462
0.25 39770 64.565 2567761 243302
0.25 43440 48.047 2087153 50662
0.25 38994 82.158 3203667 239145
0.5 45724 105.339 4770813 286980
0.5 37690 83.846 3160168 251141
0.5 38724 73.079 2829899 54829
1.00 41200 105.656 4353036 288678
1.00 43584 113.023 4926005 335726
1.00 41404 105.037 4348994 279375
2.00 40024 135.205 5411434 326434
2.00 40968 103.250 4229944 260408
2.00 40111 129.871 5209254 425061
5.00 40950 132.782 5437418 393246
5.00 40468 147.876 5984250 352233
5.00 39796 140.267 5582067 341520



Fitting a Straight Line

Description of image



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