IGEM:MIT/2006/Notebook/2006-7-31: Difference between revisions

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#find a new glycerol storage box and transfer 7/29's new glycerols into it '''-done'''
#find a new glycerol storage box and transfer 7/29's new glycerols into it '''-done'''
#nanodrop 23 minipreps from 7/29 and prepare 46 small tubes for sequencing order '''-done'''
#nanodrop 23 minipreps from 7/29 and prepare 46 small tubes for sequencing order '''-done'''
#*sequence results from tubes 11/12, 23/24, 27/28 can be discarded b/c we do not have enough dna
#troubleshoot pchBA (w/out silent mutation) PCR
#digestions+ligations+transformations (design a plan)
#digestions+ligations+transformations (design a plan)
#troubleshoot pchBA (w/out silent mutation) PCR
#get ready to fax offices about p. shuttle vectors  
#fax offices about p. shuttle vectors
#link our system device diagrams on main wiki page
#link our system device diagrams on main wiki page


Revision as of 11:42, 31 July 2006

to do

  1. find a new glycerol storage box and transfer 7/29's new glycerols into it -done
  2. nanodrop 23 minipreps from 7/29 and prepare 46 small tubes for sequencing order -done
    • sequence results from tubes 11/12, 23/24, 27/28 can be discarded b/c we do not have enough dna
  3. troubleshoot pchBA (w/out silent mutation) PCR
  4. digestions+ligations+transformations (design a plan)
  5. get ready to fax offices about p. shuttle vectors
  6. link our system device diagrams on main wiki page

Current products

  1. RBS-CDSBSMT-Term
  2. RBS-ATF1mut
  3. SPP-RBS-GFP-Term
  4. ATF1mut-Term

Digestions

  1. RBS30, 32-CDS-Term: XP
  2. RBS30, 32-ATF1: ES
  3. osmY: SP
  4. ATF1+Term: XP
  5. R0040: SP
  6. E0840: XP

Digests needed for ligation in our possession

  1. B0015: EX
  2. B0030: SP
  3. B0032: SP

Ligations

  1. R0040+1->Prom-RBS-BSMT-Term
  2. 2+Term->RBS-ATF1mut-Term
  3. RBS+4->RBS-ATF1mut-Term
  4. R0040+E0840 CP-RBS-GFP-Term
  5. osmY+RBS-SAMT-Term

pchBA PCR

Ran pchBA PCR with gradient

Sequencing Results

Looked at R40 + B30/32 sequencing results. Seems that the promoter is missing and we will be going with different set of ligations.

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