IGEM:MIT/2006/Notebook/2006-7-31

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Contents

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 23/24, 27/28 have been 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 -done, need Isadora
  6. link our system device diagrams on main wiki page -done

Current products (sequence results to verify will come in on wed 8/2)

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

Digestions (and gel extractions)

  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

Gel Extraction

  1. RBS-CDS-Term: XP 30ABC came out
  2. E0840: XP also came out
  3. RBS-ATF1: ES30/32 AB

pchBA PCR

Ran pchBA PCR with gradient -- unsuccessful again. We need advisor help.

BAT2 and THI3 PCR (and gel)

  1. primers recieved, 55 degree/standard PCR program

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.

liquid cultures

  1. R0040
  2. osmY+E0840(A,B,C)
  3. ROO30+ATF1mut(A,B,C)
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