IGEM:Missouri University of Science and Technology/2009/Notebook/Cyanobacteria Mediated Filtration of Coal Flue/2014/08/31

Cyanobacteria Mediated Filtration of Coal Flue Gas

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Date: 8/31/14 People in lab: Kira Buckowing Title: PCR of GP1 and PAO1 Temp and Gel to check products Start Time: 10:45 am Purpose: To get a positive result to nosZ Protocol: LTM Ed. 2 PCR and Gels Exceptions: PCR 1 and 3 = 2 uL DNA, 2.5 uL primers; PCR 2 and 4 = 7.5 uL DNA, 2.5 uL primers Products: Well 1 2 3 4 5 6 7 8 9 - Ladder - PCR 1 - PCR2 - PCR 3 - PCR4 Products: Results: Notes: THe band looks to be the wrong size, but its the first band that has ever shown up for something with nosZ Stop Time: 3:45 pm Next: Troubleshooting - Test primers again with controls Date: 8/31/14 People in lab: Kira Buckowing Title: innoculation of DH5 Alpha stock Start Time: 4 pm Purpose: Prep for making comp cells Protocol: LTM Ed. 2 Innoculation Exceptions: Products: Sample Label Description Source Label Quantity DH5 alphs stock 8/31 DH5 alpha cells in LB broth --- 1 Results: Notes: Stored in fridge after incubation period Stop Time: 4:05 pm Next: Competent cell creation Date: 8/31/14 People in lab: Kira Buckowing Title: PCR and Gel of GP1 and PAO1 Temp with controls Start Time: 4:30 pm Purpose: Testing primers Protocol: LTM Ed. 2 PCR and Gels Exceptions: 1 and 2 - 2 uL DNA with old primers; 3 and 4 = 7.5 uL DNA with old primers; 5 and 6 = 2 uL DNA with new primers; 7 and 8 = 7.5 uL with new primers Products: Well 1 2 3 4 5 6 7 8 9 10 - Ladder PCR1 PCR2 PCR3 PCR4 PCR5 PCR6 PCR7 PCR8 Ladder Products: Results: Bands are still too short, by about 1000 base pairs Notes: Annealing temp or primers may be the issue Stop Time: 9 pm Next: More troubleshooting