Team:CMUQ/Demonstrate

iGEM CMUQ 2019

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Members Collaborations Sponsors Attributions
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Description Details Hardware Software Demonstrate Model Results Lab Notebook
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Overview Basic Parts Improved Parts Composite Parts
Human Practices
Safety Outreach
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Hardware Software Entrepreneurship Model Measurement Characterization
Judging Form
Team Members Collaborations Sponsors Attributions Project Description Details Hardware Software Demonstrate Model Results Lab Notebook Parts Overview Basic Parts Improved Parts Composite Parts Human Practices Safety Outreach Awards Hardware Software Entrepreneurship Model Measurement Characterization Judging Form
iGEM 2019 CMUQ

Demonstrate

Our engineered system is an improvement of our 2018 project. We aimed to demonstrate the validity of the idea with more lab work and due to the need of many Cas12a proteins for testing we expressed more of it. To do so, BL21 cells were inoculated and grown to mid-log phase(OD 30-40 Kletts), then the culture was induced by IPTG for 18 hours and the proteins were harvested by being centrifuged at 5000 rpm for 5 minutes. The western-blot showed that Cas12 protein was produced and successfully purified using HA column. The functionality of the Cas12a protein was demonstrated by measuring the emitted fluorescence correlated with fragmentation using a bioanalyzer. Detection of the fragmentation indicates Cas12a identification of the SNP by successfully using the designed, SNP-specific gRNA. As to measuring the fluorescence using our device, its ability to accurately measure fluorescence was verified using DNAse Alert Kit.