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Thursday, September 1
- Started to develop Listeria Innocua project component.
- Listeria CRISPR only uses cas9 tracr gene
- Project requires:
- cas9
- Duet plasmid
- Listeria T7 RSR
- Ligate all together in biobrick
- Test with protospacer
Saturday, September 3
- 3 PCR reactions using LI as template
- Samples visualized on gel
- sample appears slightly higher than expected
- some distortion and low resolution
Sunday, September 4 to Friday, September 9
- cas9 PCR products restricted overnight
Saturday, September 10
- 9/10 restriction products dephosphorylated for 2 hours
- Overnight ligation of dephosphorylated restriction products
Sunday, September 11 to Saturday, September 17
- 9/8 restriction - 9/10 ligation products transformed into NEB cells
Sunday, September 18 to Saturday, September 24
- Attempt to assemble CMR-RSR using overlapping oligos
Sunday, September 25 to Wednesday, September 28
- Restrict, ligate, and transform CMR and pRSF. 9/25 Send sample to sequence. 9/27 Sequencing error. They will re-run.
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