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Team:Cornell/Week 17
From 2011.igem.org
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September 25th - October 1st
Sunday, September 25
Monday, September 26
Morning lab work done by: Charlie Chung, Jim Mathew
- GFP-1,2,3,4 and Control cultures were left to incubate last night at 37°C in Olin 303
- Miniprep DNA purification of the cultures to isolate the plasmid with GFP+AviTag insert
- Prepared submission for sequencing
Tuesday, September 27
Afternoon lab work done by Maneesh and Claire
Coated 2 chips using standard coating protocol
Flowed our GFP biobrick + lysate through the chip
Flow Rate = 5ul/min for 20min Control- streptavidin coated chip with water flown through
Results No binding was detected We believe there was no binding because excess biotin in the lysate was not filtered out. This excess biotin prevented the biotinylated GFP from binding to the chip.
Took pictures of previous GFP lysate experiment pictures. After 4 days there was still GFP bound to the chip. During this experiment the biotin was filtered out of the lysate before the lysate was flown through the chip. The binding was concentrated at the inlet port and the first channel of the chip. This suggests that a larger amount of lysate must be flown through to coat the entire chip or that the cells must be modified to express more of the GFP biobrick.
