Team:Arizona State/Notebook/September

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
• Began preparation of pSB1C3 plasmid for BioBrick preparation

Sunday, September 25 to Wednesday, September 28

• Finalized BioBrick preparation for intended BioBrick part submission
• Restrict, ligate, and transform CMR and pRSF. 9/25 Send sample to sequence. 9/27 Sequencing error. They will re-run.
• Wiki parties all week in the lab!
• Submitted BioBricks!