![]() We left these growing overnight on plates in the incubator. We continued with our single gene constructs Mbb1 and ccMbb1, we replated these from the successful colonies so that we could generate more. We mini-prepped the colonies and we sent them off to sequencing to gather the data and check if our duplex combination transformations were correct. We were able to see growth on the plates and we created an overnight culture from 3 different colonies. Following this PCR we ran a Gibson Assembly and a transformation and plated the cells. We ran a PCR on our genes so that we could create the duplex combinations. Therefore we opted to switch with duplex genes only, they would all contain Mbb2 and the other respective gene. We received the triplex gene sequences back and they contained the sequence of the original vector. We completed and uploaded our promotional video with a funding task to the crowdfunding platform as we opened it up to the wider public. Week 25: Invited to Complete Blog Posts and Crowdfunding Video Was Madeīiotechnologie invited our team to create a series of 6 blog posts documenting our experience in iGEM as well as explaining our project. ![]() We selected 3 colonies of each gene combination and grew them in an overnight culture. We redid the gel however the bands were not in the proper location, which meant two things: either our restriction digest was not successful or our genes do not contain the triplex genes. Due to excess noise we were going to repeat it the following lab day. We miniprepped the overnight culture and then we ran a restriction digestion to visualize the bands. From this we extracted the cells and created an overnight culture. The plates were left to grow for 3 days to have a great amount of colonies for testing. They were then plated on Kanamycin plates and left to grow in the incubator. We redid the triplex gene PCR and a Gibson assembly and transformation into DH5a competent cells. Each team was given 2 other teams to peer review and then received 2 back that could then be used to make improvements on their article before submission. The journal peer-review pairs were also assigned and sent out this week. It was then left in the facility common room for other people to play and give feedback on. The card game for our Science Communication was completed and a test round was then played by our team members. Week 24: Card Game and Peer Review for Journal New working primer stock solution for other primers. Minipreps and Restriction Digest using XbaI and NdeI. ![]() Neb Hifi Assembly:Ĭompetent cell transformation and plating on agar. Linearization of pET39b+ ( further stock). Restriction digestion and gel electrophoresis Miniprep of Mbb4 and combinations of mbb2+mbb1/mbb4/ccMbb3. Monthly reports were due again and progress was shared between the subgroups, the goals for each subgroup over the summer (month of July) were also established during this meeting. Within the week they make competent cells of the bacteria, suspended and inserted DNA as well as inserted multiple genes into the same bacteria, both those that coded for bromoform and peroxide production. Week 15: G-Block Arrive and Summer Planningįinally, the long awaited G-Blocks arrived from IDT and the lab team was ready to start modifying.
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