Team:SoundBio/Lab Notebook
Lab Notebook
From July 1st to August 16, our main goal was to clone the blue light-controlled circuit. We transformed various chromoproteins such as amilCP, tsPurple, and amilGP into a pSB1C3 and mRFP into pSB1K3, pSB1C3, and pSB1A3. After the successful transformations and minipreps, we tried cloning the circuit ourselves, but after several failures, we outsourced work to BIOFAB, a plasmid cloning service, to clone our circuits for us.
7/1 Making Cryostocks and Overnights for Competent Cell Prep
7/2 Transformations
7/3 Redo Transformations and Making SOC Media
7/5 Redo Transformations
7/9 Redo Transformations, Making TSS Solution, Growing Competent Cells, amilGFP Overnight, Making SOC Media
7/9 Co-Culture Experiments
7/10 Overnights of Successful Transformations
7/11 Miniprep and Sequencing of AmilCP, tsPurple, pSB1A3, and mRFP in pSB1K3
7/12 Miniprep of Red Light Circuit Biobricks and Making More LB Broth
7/14 Miniprep of Blue Light Circuit Biobricks
7/15 Restriction Digests of Blue Light Circuit Parts
7/15 Cloning #1 - Fail
7/16 Cloning #2 - Fail
7/19 Cloning #3 - Partially Successful
7/20 PCR's
7/21 Miniprep of Blue Light Parts
7/21 PCR Purification and PCR’s - Mostly Failed
7/22 YF1 + FixJ PCR Experiments
7/24 PCR Troubleshooting YF1 + FixJ
7/24 Minipreps of YF1 + FixJ and Ligation from Cloning #3
7/25 Cloning #4
7/26 PCR Troubleshooting - Continued
8/4 Overnights for Minipreps and Transforming GFP
8/5 Minipreps of Parts for Red Light Circuit and Characterization Plasmids
8/7 Redo GFP Transformation
8/9 Transformation of HO-1 and PcyA and Miniprep of GFP
8/10 Miniprep of GFP
7/1 Making Cryostocks and Overnights for Competent Cell Prep
7/2 Transformations
7/3 Redo Transformations and Making SOC Media
7/5 Redo Transformations
7/9 Redo Transformations, Making TSS Solution, Growing Competent Cells, amilGFP Overnight, Making SOC Media
7/9 Co-Culture Experiments
7/10 Overnights of Successful Transformations
7/11 Miniprep and Sequencing of AmilCP, tsPurple, pSB1A3, and mRFP in pSB1K3
7/12 Miniprep of Red Light Circuit Biobricks and Making More LB Broth
7/14 Miniprep of Blue Light Circuit Biobricks
7/15 Restriction Digests of Blue Light Circuit Parts
7/15 Cloning #1 - Fail
7/16 Cloning #2 - Fail
7/19 Cloning #3 - Partially Successful
7/20 PCR's
7/21 Miniprep of Blue Light Parts
7/21 PCR Purification and PCR’s - Mostly Failed
7/22 YF1 + FixJ PCR Experiments
7/24 PCR Troubleshooting YF1 + FixJ
7/24 Minipreps of YF1 + FixJ and Ligation from Cloning #3
7/25 Cloning #4
7/26 PCR Troubleshooting - Continued
8/4 Overnights for Minipreps and Transforming GFP
8/5 Minipreps of Parts for Red Light Circuit and Characterization Plasmids
8/7 Redo GFP Transformation
8/9 Transformation of HO-1 and PcyA and Miniprep of GFP
8/10 Miniprep of GFP