Differential effects of inhibitors on the regeneration of flagella Assignment

Differential Effects of Inhibitors on the Regeneration of Flagella Sherry Hebert BIOL-3221L-U2020F https://cdn.bartin.edu.tr/molekuler/ 83e62d5885d4d3edffe691a67895 5a84/46.pdf Lab Procedure Overview ● Deflagellation and drug treatment ● Collection and slide preparation ● Phase microscopy and the measurement of flagella length Part A:

Deflagellation Blend 40 mL Gonium on lowest speed (LO, WHIP) Centrifuge 3 tubes with 10 mL each for 4 minutes at 3700 rpm Part A:

Drug Treatment Discard supernatant.

Resuspend in 5 mL using each of the three treatments: medium, + colchicine, + cycloheximide).

The remainder of the nondeflagellated culture will be used as a control. Part B:

Collection IMMEDIATELY collect your time 0 samples!

Add 3 drops of culture to sampling tube and mix gently with 2 drops of glutaraldehyde:

the sample is now fixed. Part B:

Slide preparation Add a single drop of culture to the slide and place a coverslip on top:

your sample is now ready to view Continue collecting samples and preparing slides every 20 minutes for 2 hours. Part C:

Measurement of Flagella Explained in detail in Appendix G.

Phase contrast microscopy allows the flagella to be seen more easily:

Nondeflagellated Deflagellated:

60 min in Medium only Part C:

Measurement of Flagella Use program to measure the total length of the flagella Data Analysis ● Average all of the individual lengths for that particular treatment and time ● Prepare a single graph comparing the length of the flagella ( µ m) and time of treatment: all 4 treatments on single graph ● Use curved connecting lines Assignment ● Is available in the Lab 5 Nexus folder ● Is due by 11:59 PM on November 19 th and is worth 1.5% of your final grade ● Includes Figure prep, and answering questions related to the theory and analysis of your data