BME100 f2015:Group15 8amL5

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Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
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OUR TEAM

Name: Renee Chapman
Name: Malaya Prado
Name: Tedi Serreti
Name: Ibrahim Almuteb
Name: Chris Jones
Name: student
Role(s)

LAB 5 WRITE-UP

PCR Reaction Report

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

  1. Set the micro-pipette to 120 µL even though there was only 100 µL of PCR product to make sure we got it all.
  2. Mix solutions
  3. Place the glass side with the rough side up.
  4. 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.
  5. 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

High1.JPG

Image of Low Calf Thymus DNA

Low.JPG

Image of Zero Calf Thymus DNA

0.JPG

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
Mr.Gloob02.jpgMr.Gloob01.jpg


Images of Our PCR Negative and Positive Controls

Postive Download 20151103 141723.jpg Negative Download 20151103 141730.jpg


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.