Week of 01/13/25 WE'RE BACK FOR 2025



Introduction

    Welcome back everybody! This week we got the lab up and running again. Our main objective was to get our cells passaged from freezeback. Additionally, we began mixing and autoclaving media so that we would have plates and broths to work with. This year we decided to use slants instead of plates for stock for the added benefit of utilizing slants over the course of three months when plates would only ever keep cells fresh for maybe two weeks. 

    This year in the world of twitch media, we are experimenting with different dyes within the agar to better visualize the movement of cells. Updates coming next week on dyes that we decide to utilize. 

Methods


Twitch Motility Assay Protocol

1) Select media (e.g. 0.75% SMA and 1% agar)

2) Prepare a sterile hood by wiping everything down with ethanol

3) Melt media down

4) Wipe down all inoculating materials and insert them into the sterile hood

5) Dispense 10 ml of media into empty petri dishes containing five pronged molds to allow for insertion sites

a. Use a sterile 10 ml pipette

6) Let plates cool

7) Remove molds

8) Use UV light for 15 minutes to ensure sterilization of plates 

9) Inoculation center of plate (directly onto the plastic)

10) Dispense 100-150 microliters of chemoattractant media into one of the insertion sites

a. Use 1% agar media

11) Incubate Plates

12) Regularly Picture


Results

The cells from eight different species were successfully brought back from freezback. These species include: D. aquaticus P81, D. deserti, D. pimensis, D. indicus, D. radiodurans, D. xinjiangensis, D. ficus and D. budaensis. They have been gram stained and inoculated onto slants. 

Discussion

The use of slants will give us the most valuable resource any scientist can gather. Time. There are always new things to discover and new paths to take. Slants will give us our mondays back. Instead of constantly making new stock plates each week, we can instead simply draw from our stash of slants as many times as possible until contamination inevitably sets in from repeated exposure. 

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