Team:DTU-Denmark/Notebook 

       <a data-toggle="collapse" href="#collapse7" aria-expanded="false" aria-controls="collapse7"> 

       </a>

Wetlab

                           <img class="lvltwo" src="T--DTU-Denmark--moleculartools.png" alt="">
Molecular Toolbox
                               <img class="lvlthree" src="T--DTU-Denmark--pex10.png" alt="">
CRISPR-Cas9 induced PEX10knockout

The assembled CRISPRyl plasmid with PEX10 protospacers was transformed in competent E. coli DH5α cells. Purification of assembled CRISPRyl plasmid with Pex10 protospacers. The assembled CRISPRyl plasmid with PEX10 protospacers was digested with AvrII to test if the PEX10 protospacer was inserted. The digestion was successful and showed no cutting by AvrII indicating correct insertion of PEX10 protospacers. First attempt of transformation with CRISPRyl plasmid with PEX10 protospacers in Yarowia lipolytica PO1f Δku70. The transformation was unsuccessful. There was growth all over the plates. 

                               <img class="lvlthree" src="T--DTU-Denmark--ura3.png" alt="">
CRISPR-Cas9 induced URA3insertion

Confirmation of successful Gibson assembly through restriction analysis. First (unsuccessful) transformation of pCRISPRyl and pCRISPRyl+sgRNAs into Yarowia lipolytica PO1f Δku70.

                           <img class="lvltwo" src="T--DTU-Denmark--products.png" alt="">
Products
                               <img class="lvlthree" src="T--DTU-Denmark--betacarotene.png" alt="">
Beta-Carotene

BioBricks BBa_K152005 (crtE,crtI,crtYB,gfp) and BBa_K530002 (crtI) were received from the registry. E. coli with BBa_K152005 could only grow without Ampicilin, so we suspect a problem with the AmpR backbone. We decided not to use this biobrick and instead only work with BBa_K530002, BBa_K530000 and BBa_K530001.

                               <img class="lvlthree" src="T--DTU-Denmark--proinsulin.png" alt="">
Proinsulin

Designing and ordering synthetic gBlock from IDT encoding TEF1, proinsulin and GFP.


                           <img class="lvltwo" src="T--DTU-Denmark--substrate.png" alt="">
Substrates

Test shows no growth on samples of wastewater from areas contaminated with leftovers of organic molecules from fossil oil. Experiments with glycerol byproduct from Emmelev biodiesel production showed to be contaminated. We investigated where the contaminations could come from and a contaminated pipet seemed to be the source.

Compute

                           <img class="lvltwo" src="T--DTU-Denmark--hardware.png" alt="">
Hardware

The library for our LCD Display is commented in Chinese. Shoutout to James for translating. Chris, our supervisor, brought us in contact with Martin and Erik, two professors from DTU Nano and Elektro. Our university is running a new project course in January where 1st semester students ferment their own cultures and build devices to monitor growth. They built a measuring chamber and a simple circuit, reading the voltage over the photodiode with a multimeter. Isn’t there a way to do all this using an Arduino? We set a meeting for next week. Reading about digital to analog conversion(DAC) and the reverse(ADC). Dimming LEDs in the office