Team:Nevada/Week 16

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Week 16: September 3 - September 7

Contents

September 4

Joe:
gel 186 shows colonies 4, 6, 7, 8 are good

September 5

Jeremiah & Chris:
Colony PCR check 12 colonies
Check on a gel (gel #187)
Samples 8 and 9 cultured in LB + Amp

September 6

Joe:
miniprep of colonies 7, 8 RFP-SBP
glycerol stock of SBP-RFP in Top10 cells
digestion of minipreps by EcoRI and PstI
Transform to BL21 cells
culture more RFP-SBP in Top10 cells
Jeremiah & Chris:
Miniprep plasmid
Colony check plasmid again to make sure it’s not a false positive
Check on gel (gel #188)
Transform this plasmid (L. Arabanose promoter with TBP+ insert) into BL21 cells
Used Amp antibiotic on plates

September 7

Joe:
Culture colony of RFP-SBP in BL21 cells
heat inactivate RFP-SBP(EcoRI/PstI) - gel 189 confirms good
Michelle:
Culture M2-BL21 colonies from LB AMP-CM to make glycerol stocks. M2-BL21 along with purified desalted SBP-LRP :::protein used to carry out Western Transfer Blot and SDS-PAGE Coomassie staining procedure. Lysis of M2-BL21 :::pellet with protein inhibitor mixed with PBS and B-Mercaptoethanol 48.7%.
Justin and Dafne:
Large scale expression of SBP-B12 protein in BL21 cells