Pipetting the DNA samples was relatively easy. The difference between the first and second stop was easy to feel. Unfortunately all the micro-centrifuge tubes did not come out to the same volumes and one we had to scrounged for some extra master mix.Our labeling was fine and it worked well.
Fluorimeter Procedure
Smart Phone Camera Settings
Type of Smartphone: I phone 5s
Flash: Off
ISO setting: 1600
White Balance: Automatic
Exposure:High
Saturation: High
Contrast:low
Camera set-up
We put the cell phone as straight up as possible in the phone cradle and moved it to a suitable length away.
Distance between the smart phone cradle and drop = 6CM
Placing Samples onto the Fluorimeter
Set the micro-pipette to 120 µL even though there was only 100 µL of PCR product to make sure we got it all.
Mix solutions
Place the glass side with the rough side up.
Then draw up 100 µL of the solution being tested with a new sterile pipetting tip and carefully start to deposit the solution by pressing to the second stop so that it makes a bubble in between the two sections of clear glass on the slide.
Dispose of the pipette tip in the proper disposal receptical
Data Collection and Analysis
Images of High, Low, and Zero Calf Thymus DNA
Image of High Calf Thymus DNA
Image of Low Calf Thymus DNA
Image of Zero Calf Thymus DNA
Calibrator Mean Values
Initial Concentration of 2X Calf Thymus DNA solution (micrograms/mL)
Final DNA concentration in SYBR Green I solution (µg/mL)
Sample Number
RAWINTDEN DROP - BACKGROUND
'
'
MEAN
Standard Deviation
Image 1
Image 2
Image 3
5
2.5
C-1
193758284
183267482
1743467
1954573
165475
2
1
C-2
10560668
9008756
90001310
10048637
18667314
1
0.5
C-3
13001852
134533932
12967433
16850730
500851
0.5
0.25
C-4
82267074
80303172
7741956
7999660
243825
0.25
0.125
C-5
53495634
7552434
5349752
648393
1456367
0
0
C-6
148562
193548593
24037534
2836473
572484
Calibration curves
Images of Our PCR Negative and Positive Controls
Postive
Negative
PCR Results: PCR concentrations solved
PCR Product TUBE LABEL
MEAN (of RAWINTDEN DROP - BACKGROUND)
"PCR Product Concentration (µg /mL)
Total Dilution
Initial PCR Product Concentration (µg /mL)
"
+
00
00
00
00
-
00
00
00
00
1\1
00
00
00
00
1\2
00
00
00
00
1\3
00
00
00
00
2\1
00
00
00
00
2\2
00
00
00
00
2\3
00
00
00
00
PCR Results: Summary
Our positive control PCR result was 00 μg/mL
Our negative control PCR result was 00 μg/mL
Observed results
Patient 73968 : For patient one the drops overall displayed little to no green light what so ever. The overall average of the three droplets was 4207126.33.
Patient 93754 :For patient two the pictures show no green in any of the droplets except for one which had a minute amount of green, probably just an error. The overall average of patient two was 4217751.431.
Conclusions
Patient 73968 :Conclusions can't be drawn because the data that was collected is not accurate at all, because mistakes were made that rendered it useless.
Patient 93754 :Conclusions can't be drawn because the data that was collected is not accurate at all, because mistakes were made that rendered it useless.