Team:UCLA/Notebook/Protein Cages/26 June 2015
Phillip's notes
Introduction: Today, the primers for PCquad should be checked by a more experienced primer designer, and amplifying and sequencing primers should also be made. The clones from Yeate’s lab will also be picked up. Lastly, preparation for the journal presentation on PCR will be done with Nithin and Tyler.
Notes from Fasih: Design amplifying primers for the gBlock itself. It says the yield you ought to get is 1 ug, so you should get 100uL at a 10ng/ul concentration, which is great for PCR, but it wouldn’t hurt to have amplfying primers ready for emergencies Check the secondary structure on Benchling to avoid primer dimer or loops.
The cage was picked up from Dr. Yeates. The BL21 (DE3) strain is ampicillin resistant.
Designing of primers: Some notes on primer design: http://www.premierbiosoft.com/tech_notes/PCR_Primer_Design.html Potentially do two rounds of PCR to add the ends we need. Vinson suggested to use q5 polymerase Removed atg at start of PCquad
Conclusions: Due to the seemingly extensive amount of secondary structure, the primers will be worked on next week, with some feedback. Ideally, the order will go out some time mid next week.