The pre-lab write up was very helpful in getting the information for the lab ahead of time to make sure that the lab made sense and would be quicker to carry out later on. Yes, the first step involves sucking up the liquid with the pipettor, you have to push the button down and release once the tip is in the bottom of the liquid. The second step involves releasing the liquid from the pipettor, you have to push the button down as far as it goes to insure that all of the liquid was released. The final reactions had about the same amount of liquid as when they were placed in the machine. When taking the liquid from the tubes, the pipettor was placed just above 50 microliters to ensure that all of the liquid was removed. Our labeling scheme worked for the procedure, we made sure to label what number trial it was and what sample within both trials so they could be easily identified.
Fluorimeter Procedure
Imaging set-up
First the phone being used was placed into the holder making sure it was propped up to stand straight. Then the sample holder was raised using a stand to make the sample parallel to the camera lens for accurate pictures. Finally, the box was placed on top of that making sure that it was facing the right way so that the flap on the outside could be easily opened when changing samples and easily closed when taking pictures.
Placing Samples onto the Fluorimeter
Once the samples from the PCR machine were correctly diluted with solution and different concentrations, the samples were ready to be placed in the machine.
The slide that will hold the samples is placed in the slot on the machine so that the light is shining directly in between the first two rows of indentations.
The first bubble placed on the slide is 50 microliters of the first diluted solution. This was followed with 50 microliters of the green fluorescent placed on the same bubble to create one drop on the slide.
The slide was situated to make sure it was in a direct line from the light, then the timer on the phone was set and the flap was closed to make sure the inside is completely dark when the picture takes (repeat two more times to get 3 pictures total).
These steps were then repeated with the remaining concentrations of DNA samples.
Our positive control PCR result was ___-4.54271767_ μg/mL
Our negative control PCR result was __-11.65973367__ μg/mL
Observed results
Patient ___35518__ : Obtained results of -9.44915μg/mL, -3.94681067μg/mL and -10.16252367μg/mL.
Patient ___69697__ : Obtained results of -12.377727μg/mL, -15.67587233μg/mL and -12.772961μg/mL.
Conclusions
Patient __35518___ : This patient's results were inconclusive, leading to the conclusion that the patient's DNA is unlike both the positive control or the negative control.
Patient ___69697__ : This patient's results were closest to the negative result, leading to the conclusion that the patient's DNA is negative.
BME 100 Spring 2018
