Spin-concentrated all protein samples to 5-10 mg/mL
Resuspended and sonicated cell pellets
Used mosquito to set up Wizard and PEG crystal screen for eGFP-RiAFP and RiAFP
Tuesday:
Meeting with Prof. Isaacs about specific MAGE oligo designs and iGEM updates
Obtained plates of EcNR2 strain and Spec/Kan antibiotic plasmids
Wednesday:
Innoculated ECNr2 strains and prepared glycerol stocks
Designed primers for cross-over PCR to integrate RiAFP and T7 cassette into EcNR2 genome pre-MAGE
Thursday:
Found successful fuzzy-ball and shower crystals! (see image:)
Ran Lambda DE3 kit on EcNR2 strain as alternative means of integrating T7 polymerase into genome
Set up PCR amplification of pre-crossover fragments of Spectinomycin, Kanamycin cassettes and RiAFP, eGFP-RiAFP, and T7 polymerase, with ends that correspond to high-recombination frequency sites
"
