04.08.2012

GUVs meeting Experiment schedule sketch:

• LUVS and GUVs with or without Mg
• LUVS and GUVs with col oligos
• GUVs had red and green fluorescence. LUVs are dots, possible fluorescence contamination.
• Check GUVs with a normal microscope Last experiment:
• Electroformation
• Oligo coupling
• After 3 days all the GUVs and LUVs had clustered now matter is there was oligos or not. Maybe there was a problem with the linker.

Start on Monday:

• Microscope shifts begin at 9:30am. Usually are booked till 8:00pm.
• IMPORTANT: The cholesterol adding step to GUVs has to be done overnight.
• Dominik said that entropy of the system might destabilize the structure.
• Book the microscope overday and overnight
• Int 4’s without cholesterol are gonna be tested with linker and only GUVs. Linker and labeled oligos at the same time.
• How much time should the oligos be left for hybridizing? It’s around 25 mins, but we could let them for

Experiments
Experiment 1:

	LUVs+GUVs –Mg 2+	LUVs+GUVs –Mg 2+
LUVs+GUVs –Mg 2+ int4 int30 linker	LUVs+GUVs +Mg 2+ int4 int30 linker	LUVs+GUVs –Mg 2+ int4 int30

Experiment 2:

LUVs+GUVs –Mg 2+	LUVs+GUVs +Mg 2+	LUVs –Mg 2+	LUVs -Mg 2+ int4
LUVs+GUVs –Mg 2+ int4 int30	LUVs+GUVs +Mg 2+ int4 int30	LUVs+GUVs +Mg 2+ int4

Experiment 3:

Alexa int 4/linker/int 30/GUVs	Alexa int 4/linker/int 30/GUVs	LUVs/int 4/Linker	LUVs/int 4
LUVs	GUVs/int 30/Linker/LUVs/int 4/-Mg+2	GUVs/int 30/LUVs/int 4/-Mg+2	GUVs/LUVs/-Mg+2
GUVs/LUVs/int4	GUVs/int 30/Linker/LUVs/int 4/+Mg+2	GUVs/int 30/LUVs/int4/+Mg+2	GUVs/LUVs/+Mg+2

• Microscopes: 510(noon) and 780(afternoon)
• Booking System

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