IGEM:MIT/2005/Inventory

Strains
E.coli Strain Genotypes
 * 1) NB115 (pCT-4-4-20) -- 7.02 -80°C
 * 2) NB116 (pCT302 S101A) -- 7.02 -80°C
 * 3) NB118 (pCT-4M5.3) -- 7.02 -80°C
 * 4) MC4100
 * 5) *Genotype: F- araD139 delta(argF-lac)U169 prsL150 relA1 deoC1 rbsR fthD5301 fruA25 lambda- [s]

Plasmids
../Plasmid Info/
 * 1) pSB1AK3-1 -- as plate, to be made into glycerol stock thursday
 * 2) pSB1AC3-1 -- ditto
 * 3) pSB1AT3-1 -- ditto
 * 4) pSB2K3-1  -- ditto
 * 5) [[Image:PCT302.pdf|pCT302 map]]
 * 6) ../PCT302 (aka pCT-4-4-20)/-- 7.02 4°C
 * 7) ../PCT302 S101A/ -- 7.02 4°C
 * 8) ../PCT-4M5.3/ -- 7.02 4°C

PCR Amplification

 * 1) Spin PCR purification kit
 * 2) Platinum, TAQ polymerase (DNA polymerase for PCR)
 * 3) DNTP mix (25mm ea. dntp) (nucleotides for PCR)

Agarose gels

 * 1) Agarose 100g	(pouring gels)
 * 2) 10X TAE buffer (pouring/running gels)
 * 3) 1 kb DNA ladder 250µg (molecular weight standard)
 * 4) Ethidium Bromide Solution (10mg/mL) (staining DNA)

Cutting/joining DNA fragments

 * 1) EcoR1 10,000 u (Restriction Enzyme)
 * 2) PstI 10,000 u (Restriction Enzyme)
 * 3) SpeI 2500 u	(Restriction Enzyme)
 * 4) Xba1 3,000 u	(Restriction Enzyme)
 * 5) T4 DNA Ligase 100,000un (Ligating DNA fragments)

Purifying DNA

 * 1) Spin Plasmid Kit, Mini(for minipreps)
 * 2) Gel Extraction Kit 50 (purifying from gel slices)
 * 3) Spin PCR Purification Kit (purifying PCR products)

Microbiology supplies

 * 1) LB amp -- Cold Room
 * 2) Amp stock -- -20 Fridge
 * 3) VWR Petri Dish (100x15mm, case of 500) (for pouring plates)
 * 4) Test Tube, Culture (Polystyrene, 12x75, case of 1000)	(special tubes for FACS)

Bio Bricks
pSB1AT3-1 pSB1AC3 pSB1AK3
 * 1) BBa_J07012 scFv anti-fluorescein [4-4-20]
 * 2) BBa_J07013 scFv anti-fluorescein [S101A]
 * 3) BBa_J07014 scFv anti-fluorescein [4M5.3]
 * 4) BBa_J07017 FecA protein (full length)
 * 1) BBa_J07021 FecR
 * 2) BBa_J07022 FecI
 * 1) BBa_J07043 J07012.B0015 scFv4-4-20.TT
 * 2) BBa_J07044 J07013.B0015 scFvS101A.TT
 * 3) BBa_J07045 J07014.B0015 scFv4M5.3.TT

Miniprepped DNA

 * 1) R0053 -- Amp -- KBlab -- 4°C
 * 2) R0040 -- Amp -- KBlab -- 4°C
 * 3) R0011 -- Amp -- KBlab -- 4°C
 * 4) ../Quad part inverters/
 * 5) E0420 -- Amp -- KBlab -- 4°C
 * 6) E0430 -- Amp -- KBlab -- 4°C
 * 7) E0840 (rbs:gfp:term)
 * 8) B0015 (term)
 * 9) R0040 (tetR promoter)
 * 10) R0011 (lacI regulated promoter)
 * 11) B0030 (Strong RBS)
 * 12) J13002 (R0040.B0034)

Streaked out on a plate

 * 1) I0500 (pBad/AraC promoter)

Ordered as Stabs

 * 1) R0010
 * 2) R0051
 * 3) J04500
 * 4) I1030
 * 5) I1031
 * 6) I1032
 * 7) I1033

Assemblies Ordered from Registry
note: the registry probably will not make these in any reasonable time frame. We need to make them ourselves.
 * 1) J07036
 * 2) J07037
 * 3) J07038
 * 4) J07039
 * 5) J07040
 * 6) J07041
 * 7) J07042

Planning/Building Bio Bricks

 * 1) BBa_J07005 [msr] E.coli JM83 alkaline phosphatase gene (PhoA) 1416 bp
 * 2) BBa_J07006 [tmp] malE  1191 bp
 * 3) BBa_J07007 [reg] ctx promoter  145 bp
 * 4) BBa_J07008 [tmp] ToxR w.t. (from Vibrio Cholarae, aa's 1-294)  1160 bp
 * 5) BBa_J07009 [reg] ToxR' (aa's 1-210)  630 bp
 * 6) BBa_J07010 [reg] ToxR inner (aa's 1-198; cytoplasm + TM) 594 bp
 * 7) BBa_J07011 [rpt] ctx promoter :: gfp reporter	 871 bp

Specified:
Please click link in order to see ../Sequences of primers specified/. They consist of the following:

Ordered/Received:
Arrived: 8/4/2005 fwd & rev138 fwd & rev155 fwd & rev176 fwd & rev178 fwd180 fwd & rev527 fwd & rev571 fwd & rev138-141 fwd & rev155-158 fwd & rev176-180 fwd & rev178-182 fwd & rev180-184 fwd & rev521-529 fwd & rev523-531 fwd & rev527-531 fwd & rev567-576 fwd571-574 Arrived: 8/5/2005 Rev180 Rev571-574
 * Not completely or not received primers are listed in italics
 * 1) scFv fwd, rev
 * 2) FecA fwd, rev, mid-fwd, mid-rev
 * 3) FecA promoter fwd, rev
 * 4) MalE fwd, rev
 * 5) PhoA fwd, rev, mid1-fwd, mid1-rev, mid2-fwd, mid2-rev
 * 6) FecI fwd, rev
 * 7) FecR fwd, rev, mid-fwd, mid-rev
 * 8) ToxR ATCC fwd, rev
 * 9) Linker fwd1, rev1, fwd2, rev2, rev3,
 * 10) scFv fwd flag test (for introducing a start codon)
 * 11) FecA knockout with Tetracycline marker
 * 12) Fur knockout with Chloramphenicol marker
 * 13) FecA promoter Reverse, version2
 * 14) Fusion

(n.b: "mid" refers to primers intended to introduce point mutations to take out restriction sites in our parts)