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.
Smart Phone Camera Settings
- Type of Smartphone: I phone 5s
- Flash: Off
- ISO setting: 1600
- White Balance: Automatic
- Saturation: High
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)
||RAWINTDEN DROP - BACKGROUND
| ||Image 1||Image 2||Image 3||
Images of Our PCR Negative and Positive Controls
PCR Results: PCR concentrations solved
|PCR Product TUBE LABEL
||MEAN (of RAWINTDEN DROP - BACKGROUND)
||"PCR Product Concentration (µg /mL)
||Initial PCR Product Concentration (µg /mL)
PCR Results: Summary
- Our positive control PCR result was 00 μg/mL
- Our negative control PCR result was 00 μg/mL
- 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.
- 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.