Team:Nevada/Week 16

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<center><font size=4><b>Week 16: September 3 - September 7</font size></b></center>
<center><font size=4><b>Week 16: September 3 - September 7</font size></b></center>
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==September 3==
 
==September 4==
==September 4==
 +
:Joe:
 +
:::gel 186 shows colonies 4, 6, 7, 8 are good
==September 5==
==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==
==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==
==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

Revision as of 03:51, 4 October 2012



Protocols | Week 1 | Week 2 | Week 3 | Week 4 | Week 5 | Week 6 | Week 7 | Week 8 | Week 9 | Week 10 | Week 11 | Week 12 | Week 13 | Week 14 | Week 15 | Week 16 | Week 17 | Week 18 | Week 19 | Week 20 | Week 21


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