Hi all, Attached is the much awaited gel picture of the plasmid preps. It is in the order A to F, 1 to 4(i.e. A1, A2..

A4, C1, C2.... etc.) from left to right. You should bear in mind the spot you were working at on that day to figure out your particular prep.The samples were run along with Lambda Hind III marker DNA (picture and bands are provided in the lab manual), on a 1% agarose gel. Those of you who guested need to e-mail your host TAs to send you the gel pictures. Those of you who could not make it for the lab can interpret and comment generally on the size of the expected band(s), their quality etc. Also attached is a sample lab report on the Lab math class. This is just a guide to give you a sort of skeleton/idea on which you can build/base your lab report. For the 'results' and 'discussion' sections, don't restrict yourself to the post-lab questions in the lab manual. The post- lab questions are also a guide to help you formulate the 'results' section and the 'discussion' section. The gel pictures for the restriction digestion lab will be sent in another e-mail, shortly. Good luck! and see you next week!

Lab Math, Oh How Sweet it is?!

Pre-lab:
In this activity, we will learn the concepts of concentration and its uses in a laboratory as well as its relevance to most biological systems. The techniques covered in this lab will be how to measure and formulate solutions at a prescribed concentration. This activity will require that a tea solution of a prescribed concentration be made by a pair of students. All tea solutions will then be tasted blindly and the result(s) of each tasting will be recorded.

Materials & Methods:

Each step in the activity was performed in accordance to Dr. Pattisons BCHS3201 protocols. Below are the recorded deviations: A 200 mL solution was made instead of 150 mL. A 100 mL graduate cylinder was provided for making solution. Since this instrument cannot measure the required volume of 200 mL, the sugar was first dissolved in 50 mL of tea in a cup. This volume was noted and the remainder volume needed to make a 200 mL solution was added to the existing 50 mL. The required sugar was calculated as follows: Example:

Results:
Table I: Tabulation of the sugar tasted in the blind tasting of tea samples from both Tuesdays and Wednesdays samplings. Sample 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 Actual Sugar Concentration (M) 0.01 0.001 0.2 0.005 0.02 0.04 0.0001 0.08 0.1 0.15 0.5 0.005 0.3 0.15 0.4 0.2 0.25 0.08 0.1 0.02 (+) Response 0 2 18 1 11 18 0 18 18 18 17 0 17 17 17 17 17 9 14 0

Plot I: Responses to sugar sensing plotted with semi-logarithmic scale on the x-axis. Anomalous tasting of sugar below the human sugar sensing threshold of 0.01 M was observed.

(+) Response to Sugar in Tea

20 16 12 8 4 0 0.0001

Aggregate response to Sugar from Tuesday Samplings

0.04 0.08 0.1 0.02 0.15 0.2

0.005

0.01

0.001

0.01 [Sugar] (M)

Wednesday
0.1

Tuesday

Plot II: Responses to sugar sensing plotted with semi-logarithmic scale on the x-axis. Sugar sensing seemed to be not as sensitive in this group since sweetness should have been detected at 0.02 M given that the sugar sensing threshold was 0.01 M. Perhaps the groups assigned to the sugar concentration of 0.02 M did not dissolve the correct amount of sugar.

(+) Response to Sugar in Tea

20 16 12 8 4

Aggregate response to Sugar from Wednesday Samplings

0.15 0.1 0.08 0.25 0.2 0.3 0.4 0.5

0.005

0.02

0 0.0001

0.001

0.01 [Sugar] (M)

0.1

Discussion/Post-Lab Questions:
1. What is the lowest concentration at which a student in your class was able to taste sugar? The lowest concentration where participants were able to taste sugar was 0.001M for the Tuesday samplings and 0.08 M for the Wednesday samplings. 2. Did anyone claim to taste sugar when the sample was actually the negative control (tea but no sugar at all)? Why do you think this might have happened? Relate this to the placebo effect in medical practice. Blind tastings of a negative control was not done so one could not address this question. However, a few participants from the Tuesday samplings did taste sugar below the human threshold of 0.01 M. It is likely that the participants did not clean their palates well so residual sugars from the previous tasting confounded their interpretation of the tea. With respect to the placebo effect in medical practice, a positive response might have occurred given that participants were psychologically primed to detect a positive signal, i.e. the taste of sweetness. In addition, responses were collected in aggregate and not individually so the placebo effect, if occurred, would have been compounded by peer pressure. 3. At what concentration of sugar can everyone in you class taste sugar?

All participants from the Tuesday samplings were able to taste sugar at 0.04 M while participants from the Wednesday samplings reported a higher concentration of 0.15 M. 4. What was the concentration of sugar you made? Was your guess above or below the threshold of detection by the students in the class? This participant was the TA and thus exempt from this question. ;) 5. Now think about the significance of differences in sugar sensing on a broader scale. If you were a research scientist, what might the next question you ask based on this data be? The next question to ask that is relevant to sensing on a broader scale is whether the other tastes, i.e. bitter, salty, and sour, operate on the same threshold of 0.01 M. It is expected that bitter taste will have the lowest threshold since bitter tasting things in nature are usually poisonous. Thus there would have been a positive selective pressure for the keener sensing of this taste else most life forms would quickly cease to exist.