New cloning plasmid for FoxP homology domains

on Wednesday, February 14th, 2018 1:14 | by

UNIVERSITY PART

Map of the phd-ds-Red-attP plasmid used for cloning: 1 desired construct is aiming to target all dFoxP isoforms with one homolgy domain starting at exon 1 to exon 3, the second homology domain starting at exon 3 to exon 6. The second construct is aimed to target just dFoxP IsoB, with both the homology domain at the end of the protein. This plasmid is aimed to knock down the FoxP gene without the Gal-4 knock-in.

 

PCR product of the homology domains 1 and 2 for FoxP (upper part of the image, left and right), and homology domains 1 and 2 for FoxP-isoformB (lower part of the picture, left and right): amplification of the desired homology domains via PCR (5 replicates for each construct in order to test different annealing temperatures). This products are going to be used for the subsequent cloning in the plasmid.

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Starting of the FoxP project

on Monday, February 5th, 2018 11:01 | by

UNIVERSITY PART

Maps of the FoxP gene and of the pTGem plasmid used for cloning: 1 desired construct is aiming to target all dFoxP isoforms with one homolgy domain starting at exon 1 to exon 3, the second homology domain starting at exon 3 to exon 6. The second construct is aimed to target just dFoxP IsoB, with both the homology domain at the end of the protein.

 

 

 

PCR product of the homology domains 1 and 2 for FoxP (upper part of the picture, left and right), and homology domains 1 and 2 for FoxP-isoformB (lower part of the picture, left and right): amplification of the desired homology domains via PCR (4 replicates for each construct in order to test different annealing temperatures). The products will be used for subsequent cloning in the vector.

 

Plasmid with Hom1 Hom2 FoxP ready: the image shows the presence of the homology domain 2 for dFoxP in a plasmid where we already ligated the homolgy domani 1. This construct is thus ready to be injected.

 

KLINIKUM PART

We transfected E.coli cells with the plasmids for IsoA and B from the Schaff lab (Berlin) and subsequently we sequenced them with 4 primers each (1 FW primer sequence in the plasmid upstream the ORF and 1 RV, 1 FW primer sequence in the middle of the ORF and 1 RV).

pcDNA plasmid IsoA correctly amplified in E.coli

 

pcDNA plasmid IsoB correctly amplified in E.coli

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quality test before strokelitude experiments

on Monday, January 29th, 2018 12:40 | by

Some traces from this week just so that you have an idea how do they look like. To me they are not the optimal traces I expected. But one can see some signal there. I will start the screen hoping to get enough good traces without too much work.

what do you think is the best quality control for accepting a trace for the analysis or not. I was thinking the 3D mapping gives a good hint but without quantification.

In addition I was writting to Andi Straw to solve the issue of running two cameras in the same computers, but now answer

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Summary of results of master thesis

on Monday, December 11th, 2017 12:35 | by

Self-learning of Rover/sitter from flight simulator

Long-term memroy from self-learning

 

Self-learning training/tests from transgeneic flies

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Chrimson/Gr28bd lines tests in yellow light (580nm)

on Monday, December 4th, 2017 2:17 | by

 

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Better analysis of slope dependence on flight length and further insights in the SMAP code

This is another way of showing how the slope of the SMAP analysis varies with length. I have choped the time series in 4 chunks and saw what was the slope for the chunks and for the whole time series. I think this is the best statistical way of doing it, it should not depend on what line was tested. I cannot see any effect.

 

Here below is just to show what I found out in the code. What I thought that theta was controlling for nonlinearity in the model, for me it seems rather a control for under- overfitting in the model. So I might need to read the paper and see what do they say, and if it is the same as what I found out in the code.

 

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Do not agree with Sathish?

on Monday, November 27th, 2017 2:43 | by

So this is the final graph assuming that 20Hz is the sampling rate. Sathish was not sure what it was and he said he will check.

 

 

 

To have a better overview how the length of the flight compares with the slope obtained from the SMAP. There is no big correlation whatsoever with around 100 flies. Sathish found this in his thesis with data from seventy something WTB, but to me it seems like an anecdotal result.

Here the same as above, just for showing the fit line.

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new TrpA/chrimson self-learning tests

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NorpA; Chrimson/Gr28 (N=12, ATR)

NorpA; Chrimson/Gr28 (N012, No ATR)

NorpA;Chrimson/+ (N=11, ATR)

 

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Thornlabs spectrometer working!

on Monday, November 20th, 2017 2:59 | by

After getting a new spectrometer, we confirmed that the first one was faulty. Comparison of old (1st and 3rd measures) and new (2nd and 4th) spectrometer. Now much more sensitive and the right spectrum measured for the green LED present in the spectrometer itself and for the light comming out of the light guide coupled with the red LED (whose spectrum does not seem to change after travelling through the light guide)

 

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All sathish data analysis

This are the results of all of the flies analyzed from Sathish. I just need to know the frequency of the acquisition to exclude the ones that are below 6 minutes.

 

 

 

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