IGEM:MIT/2005/Experiments: Difference between revisions
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==Experiment Status== | ==Experiment Status== | ||
''' | '''Stock-up MiniPreps''' | ||
[[Image:Check.gif]] | [[Image:Check.gif]] grow up B0015, J13002, E0840 | ||
[[Image:Check.gif]] | [[Image:Check.gif]] overnight B0015, J13002, E0840 | ||
[[Image:Check.gif]] | [[Image:Check.gif]] miniprep B0015, J13002, E0840 | ||
[[Image:Check.gif]] | [[Image:Check.gif]] glycerol stock J13002 | ||
'''FecA (full length)''' | '''FecA (full length)''' | ||
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[[Image:Check.gif]] ligate together | [[Image:Check.gif]] ligate together | ||
[[Image:Check.gif]] transform | [[Image:Check.gif]] transform | ||
[[Image: | [[Image:Check.gif]] overnights for miniprep | ||
[[Image: | [[Image:Check.gif]] identify correct clones | ||
[[Image: | [[Image:Check.gif]] overnights for freezer stock | ||
[[Image:Check.gif]] freezer stocks | |||
===Jessica=== | ===Jessica=== | ||
'''MalE''' | '''MalE''' | ||
[[Image:Check.gif]] PCR | [[Image:Check.gif]] PCR | ||
[[Image: | [[Image:Check.gif]] digest MalE + pSB1AK3-1 plasmid w/ E/S | ||
[[Image: | [[Image:Check.gif]] gel purify each | ||
[[Image: | [[Image:Check.gif]] ligate together | ||
[[Image:Pending.gif]] transform | [[Image:Pending.gif]] transform | ||
[[Image:Pending.gif]] identify correct clones | [[Image:Pending.gif]] identify correct clones | ||
'''PhoA''' | '''PhoA''' | ||
[[Image: | [[Image:Check.gif]] PCR with higher annealing temperature | ||
[[Image: | [[Image:Check.gif]] digest PhoA+ pSB1AK3-1 plasmid w/ E/S | ||
[[Image: | [[Image:Check.gif]] gel purify each | ||
[[Image: | [[Image:Check.gif]] ligate together | ||
[[Image:Pending.gif]] transform | [[Image:Pending.gif]] transform | ||
[[Image:Pending.gif]] identify correct clones | [[Image:Pending.gif]] identify correct clones | ||
'''PhoA'''' | |||
[[Image:Pending.gif]] QuikChange | |||
===Annie=== | ===Annie=== | ||
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[[Image:Pending.gif]] ligate together | [[Image:Pending.gif]] ligate together | ||
[[Image:Pending.gif]] transform | [[Image:Pending.gif]] transform | ||
[[Image:Pending.gif]] identify correct clones | [[Image:Pending.gif]] identify correct clones (digest) | ||
[[Image:Pending.gif]] freezer stock | |||
'''FecI''' | '''FecI''' | ||
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[[Image:Check.gif]] digest FecI + pSB1AK3-1 plasmid w/ E/S | [[Image:Check.gif]] digest FecI + pSB1AK3-1 plasmid w/ E/S | ||
[[Image:Check.gif]] gel purify each | [[Image:Check.gif]] gel purify each | ||
[[Image: | [[Image:Check.gif]] ligate together | ||
[[Image: | [[Image:Check.gif]] transform | ||
[[Image:Pending.gif]] identify correct clones | [[Image:Pending.gif]] identify correct clones (digest) | ||
[[Image:Pending.gif]] freezer stock | |||
'''FecR''' | '''FecR''' | ||
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[[Image:Check.gif]] digest FecR + pSB1AK3-1 plasmid w/ E/S | [[Image:Check.gif]] digest FecR + pSB1AK3-1 plasmid w/ E/S | ||
[[Image:Check.gif]] gel purify each | [[Image:Check.gif]] gel purify each | ||
[[Image: | [[Image:Check.gif]] ligate together | ||
[[Image: | [[Image:Check.gif]] transform | ||
[[Image:Pending.gif]] identify correct clones | [[Image:Pending.gif]] identify correct clones (digest) | ||
[[Image:Pending.gif]] freezer stock | |||
'''FecA'''' | |||
[[Image:Pending.gif]] QuikChange | |||
'''FecR'''' | |||
[[Image:Pending.gif]] QuikChange | |||
==Project Experiments== | ==Project Experiments== | ||
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'''Actuator''': [[../Actuator#Experiments/|Exp. Here]] | '''Actuator''': [[../Actuator#Experiments/|Exp. Here]] | ||
[[../Experimental Timeline/]] | |||
Latest revision as of 22:34, 14 August 2005
[[../Experimental Flowchart/]]
indicates done! indicates pending...
Experiment Status
Stock-up MiniPreps grow up B0015, J13002, E0840 overnight B0015, J13002, E0840 miniprep B0015, J13002, E0840 glycerol stock J13002
FecA (full length) PCR bb ends digest FecA + pSB1AK3-1 plasmid w/ E/S gel purify each ligate together transform overnights for miniprep identify correct clones overnights for freezer stock freezer stocks
Jessica
MalE PCR digest MalE + pSB1AK3-1 plasmid w/ E/S gel purify each ligate together transform identify correct clones
PhoA PCR with higher annealing temperature digest PhoA+ pSB1AK3-1 plasmid w/ E/S gel purify each ligate together transform identify correct clones PhoA' QuikChange
Annie
FecA promoter PCR with current primers with higher annealing temperature PCR with newly designed primers digest FecApro + pSB1AK3-1 plasmid w/ E/S gel purify each ligate together transform identify correct clones (digest) freezer stock
FecI PCR bb end digest FecI + pSB1AK3-1 plasmid w/ E/S gel purify each ligate together transform identify correct clones (digest) freezer stock
FecR PCR bb end digest FecR + pSB1AK3-1 plasmid w/ E/S gel purify each ligate together transform identify correct clones (digest) freezer stock
FecA'
QuikChange
FecR'
QuikChange
Project Experiments
Input Device: [[../Input: Ligand#Experiments/|fluorescein]] Head Receiver Unit: [[../Receiver Head Unit: scFv#Experiments/|anti-fluorescein scFv]] Receiver 1: [[../Receiver 1: ToxR#Experiments/|ToxR]] Receiver 2: [[../Sensor 2#Experiments/|FecA]] Signal Processor: [[../Signal processing#Experiments/|Exp. Here]] Actuator: [[../Actuator#Experiments/|Exp. Here]]
[[../Experimental Timeline/]]