Control of general FoxP expression patter (line 104y)

Expression of 104y (green) and FoxP (red)

There seems to be no colocalisation of 104y and FoxP.

Overlap of 121y GFP expression with FoxP expression is the fan-shape body.

Problems with line c205, no GFP expression.

A,B: KO in embyro stage
C: KO in larvae stage (Experiment 1: in 30 °C for 18 hours, Experiment 2: in 30 °C for 36 hours)

We phenocopied the results of the ELAV-Gal4 cross.

Now we are waiting for the results of the Temperature switch at the second instar larvae stage

Knockout of FoxP in adult Drosophila, using nsyb Gal4 gen-switch line 80699. Flies were kept for two days on RU486 food.

PCR pCDF6 Primer
-12.5 µl primer reverse/forward
-5 µl dNTPs
-2.5 µl Polymerase
-0.57 µl pCDF6 undigested
-50 µl Q5 buffer
-166.93 µl H2O
Agarose-gel electrophoresis
E.Z.N.A. Gel Extraction

pCDF6 NEBuilder Assembly Reaction
-pcr1: 4.85 ng -> 0.88 µl
-pcr2: 4.24 ng -> 1.5 µl
-pcr3: 4.87 ng -> 1.6 µl
-pCDF6 digested: 100 ng -> 2.8 µl
-H2O: 3.22 µl
-10 µl NEBuilder HiFi DNA Assembly Master Mix/Control
PCR
heat shock transformation of the construct into competent E.coli cells
-> plated on LB0+Amp plates
-> no colonies

primer: reverse/forward each 10 µl
dNTPs: 4 µl
buffer: 40 µl
polymerase: 2 µl
H2O: 133.6 µl
undigested vector: 0.4 µl