Success: rsh Stock has rsh1 Mutation

on Monday, August 7th, 2023 11:11 | by

Via gDNA analysis and PCR was the specific area of the rsh gene extracted and amplified where the nucleotide substitution: C to T (Folkers et al., 2006) should be for the rsh1 mutation. The amplicon was Sanger sequenced which proved the nucleotide substitution.

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Bachelor Blog / #3 finally, data!

on Monday, July 24th, 2023 2:22 | by

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Little by little

on Monday, July 24th, 2023 1:59 | by

Despite very warm weather, some flies did fly, even though the learning performance of the control flies was really poor. At least for now, it looks like all stocks are learning and that rut and rsh flies learn at least equally well as the Berlin flies. I’ve also managed to fix the positive preference problem:

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Pure vs Sugar Preference results

on Monday, July 24th, 2023 9:24 | by

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Creating gRNAs via PCR

on Friday, July 7th, 2023 6:38 | by

1% Agarose gel with 100bp marker and PCR1 rsh, PCR2 rsh and PCR3 rsh or PCR1 rut, PCR2 rut and PCR3 rut, respectively.
The template pCFD6 was used with a concentration of 640 pg/µl.
1% Agarose gel with 100bp marker and PCR1 rsh, PCR2 rsh and PCR3 rsh or PCR1 rut, PCR2 rut and PCR3 rut, respectively.
The template pCFD6 was used with a concentration of 64 pg/µl (1:10 dilution).
1% Agarose gel with 100bp marker and PCR1 rsh, PCR2 rsh and PCR3 rsh or PCR1 rut, PCR2 rut and PCR3 rut, respectively.
The template pCFD6 was used with a concentration of 64 pg/µl (1:10 dilution).
50µl of 5xQ5 High GC Enhancer was added to the PCR mix.
1% Agarose gel with 100bp marker and PCR1 rsh, PCR1 rut.
The template pCFD6 was used with a concentration of 128 pg/µl (1:5 dilution).
50µl of 5xQ5 High GC Enhancer was added to the PCR mix.
1% Agarose gel with 100bp marker and PCR1 rsh, PCR2 rsh and PCR3 rsh or PCR1 rut, PCR2 rut and PCR3 rut, respectively.
The template pCFD6 was used with a concentration of 640 pg/µl.
The Phusion DNA Polymerase was used instead of the Q5 High-Fidelity DNA Polymerase.
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Pure vs salt preference results

on Friday, July 7th, 2023 12:57 | by

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Kicked out Canton S

on Thursday, June 29th, 2023 5:25 | by

Since the Canton S strain I used wasn’t a perfect genetic background strain anyway and didn’t fly properly, I kicked it out and replaced it with the wild type Berlin data I had collected just prior to the rut and rsh mutants. Now I can collect data for two projects in one go: I check the rut/rsh learning mutants if they still behave the way they should and with the wtb control strain and continue collecting data to evaluate their optomotor responses after training. So now the learning scores for the three strains look like this:

It still doesn’t look like rut is better than wtb, so I’m still skeptical that the strain is really what it should be. The rsh data also don’t look very promising, but I still need to get more flies with a negative preference before the training.

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Suspicion and progress!

on Monday, June 26th, 2023 2:03 | by

Suspicion: rut may not be rut

Carsten Duch is identifying the muscles involved in operant self-learning:

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Preference results: pure vs salt

on Monday, June 26th, 2023 1:45 | by

The first plot of each group show my results from the week before, and the ones beneath show the final results for red and blue light, after filling the sample size up to 20n.

Red light:

for comparison, this plot shows the control groups for the remaining 8n I had left separately:

Blue light:

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Preference Results for Canton S

on Monday, June 19th, 2023 1:20 | by

The following shows the preference and avoidance towards Sugar and Salt for the Canton S larvae. In addition, the larvae were tested unter blue/room light, to see if there were any differences.

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