Harvard BioDesign/24 July 2014

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Picked colonies from July 23rd transformations to grow up Place in 3 milliliters of LB + carb Picked 11 colonies to grow

Add 7 milliliters of LB+ carb to the already growing transformations from yesterday Though it took a while for the Mach I cells to grow, it seems like they grew a lot overnight, so we wanted to grow up more → more plasmid product 3 colonies currently growing up

Ran PCR on primer and backbones to prepare for another Gibson reaction to put the insert into the pBbE1a backbone with SpyTag and CsgA The Mach I cells with the plasmid seem to be having a lot of difficulty growing up on the plates and in the culture tubes We went one step back by retransforming The plates from this were still very weak, and we will find out after sequencing if in fact the plasmid was inserted correctly Now we are going one more step back, to the start of it all→ the Gibson reaction

Joshi Meeting

Sherwin Williams Explained our altered direction with them They are very interested Want to talk about a more formal collaboration with the project Mentioned James- microrheology Mentioned Connie- microfluidics Work on Perhaps making it a top coat Manufacturing moisture so that E. coli stay alive Having them stay moisturized for a day is going to be a challenge A substance that absorbs lots of water and then can release it slowly Polyacrylics Alginate

This week Should have SpyTag and CsgA by the end of today

Tracks are chosen online

Meeting with Johnathan to talk about our wiki Michelle and Nicholas have been working on possible design ideas

Testing through Gel shift His column Curli purification comparison

We are interested in writing an abstract for the BMES conference, but we are hesitant because we don’t actually have results

Pull down assay → replace the gel shift

Congo red → make sure you’re still getting curli