IGEM:MIT/2007/Notebook/2007-8-1: Difference between revisions
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#Decide on constitutive promoter for CPX | #Decide on constitutive promoter for CPX | ||
#*Transform promoter into DH5a | #*Transform promoter into DH5a | ||
#*Plate transformants | |||
#Check sequencing results for Mer/GFP 3A | #Check sequencing results for Mer/GFP 3A | ||
#Perform flourescence assay on mer-gfp construct | #Perform flourescence assay on mer-gfp construct | ||
#Clean out fridge | |||
===Flourescence Assay Protocol=== | ===Flourescence Assay Protocol=== | ||
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*mer.I13500 in pSB1AT3 came back really crappy. Biobrick prefix and suffix are partially present, cannot locate the sequence for GFP. | *mer.I13500 in pSB1AT3 came back really crappy. Biobrick prefix and suffix are partially present, cannot locate the sequence for GFP. | ||
*Need to send in the parts for sequencing. | *Need to send in the parts for sequencing. | ||
*Will run a colony PCR as a diagnostic. | *Will run a colony PCR as a diagnostic? | ||
===Constitutive Promoter=== | |||
Construct containing CPX and the polystyrene-binding peptide that will hopefully go into final system: | |||
I14032/R0051.B0034.CPX-insert. | |||
I14032 is a lacI repressable constitutive promoter. R0051 is a lambda cl repressable promoter. | |||
===Things That Got Cleaned Out=== | |||
Ask Semmie. | |||
Latest revision as of 14:24, 2 August 2007
Agenda
- Decide on constitutive promoter for CPX
- Transform promoter into DH5a
- Plate transformants
- Check sequencing results for Mer/GFP 3A
- Perform flourescence assay on mer-gfp construct
- Clean out fridge
Flourescence Assay Protocol
- Draw sample of 1.5 ml from incubated cells with Hg
- Plot OD's
- Spin down 3 mins at 13000 rpm
- Aspirate and resuspend in 1.5 ml .9% NaCl
- Repeat 3
- Repeat 4
- Put 150ul in plate reader
Sequencing Results
- mer.I13500 in pSB1AT3 came back really crappy. Biobrick prefix and suffix are partially present, cannot locate the sequence for GFP.
- Need to send in the parts for sequencing.
- Will run a colony PCR as a diagnostic?
Constitutive Promoter
Construct containing CPX and the polystyrene-binding peptide that will hopefully go into final system:
I14032/R0051.B0034.CPX-insert.
I14032 is a lacI repressable constitutive promoter. R0051 is a lambda cl repressable promoter.
Things That Got Cleaned Out
Ask Semmie.
