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

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-CDS_BSMT-Term
2. RBS-ATF1_mut
3. SPP-RBS-GFP-Term
4. ATF1_mut-Term

Digestions (and gel extractions)

1. RBS_{30, 32}-CDS-Term: XP
2. RBS_{30, 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+ATF1_mut(A,B,C)