Difference between revisions of "Part:BBa R0061:Experience"
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=====Materials & Methods===== | =====Materials & Methods===== | ||
*plamisd used | *plamisd used | ||
− | *#[[Part:BBa_K396012|K396012]] on pSB2K3 (parts which GFP is inserted | + | *#[[Part:BBa_K396012|K396012]] on pSB2K3 (parts which GFP is inserted under R0061) |
*#[[Part:BBa_K396011|K396011]] on pSB1A3 (constitutive LuxR generator) | *#[[Part:BBa_K396011|K396011]] on pSB1A3 (constitutive LuxR generator) | ||
*#[[Part:BBa_J09250|J09250]] on pSB2K3 (GFP under lac promoter) as a positive control | *#[[Part:BBa_J09250|J09250]] on pSB2K3 (GFP under lac promoter) as a positive control | ||
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**Overnight cultures were diluted to 1:100 and incubated in a fresh LB medium (containing 0 or 1 uM 3OC6HSL and antibiotics) at 37 ºC for 12 h. | **Overnight cultures were diluted to 1:100 and incubated in a fresh LB medium (containing 0 or 1 uM 3OC6HSL and antibiotics) at 37 ºC for 12 h. | ||
**OD<sub>600</sub> was measured, and 200 µL of the cultures were used to measure GFP fluorescence with a fluorescence microplate reader(excitation 485 nm,emission 527 nm). | **OD<sub>600</sub> was measured, and 200 µL of the cultures were used to measure GFP fluorescence with a fluorescence microplate reader(excitation 485 nm,emission 527 nm). | ||
+ | |||
=====Results===== | =====Results===== | ||
[[Image:Chiba R0061.png|right|300px|thumb|'''Figure'''. Fluorescence of GFP under lux repression promoter with/without AHL. p.c. stands for positive control, n.c. stands for negative control. All biobricks are co-transformed with K396011 (constitutive luxR generator). Bar heights are normalized to OD<sub>600</sub> and represent the averages of three replicates; error bars, standard deviations. ]] | [[Image:Chiba R0061.png|right|300px|thumb|'''Figure'''. Fluorescence of GFP under lux repression promoter with/without AHL. p.c. stands for positive control, n.c. stands for negative control. All biobricks are co-transformed with K396011 (constitutive luxR generator). Bar heights are normalized to OD<sub>600</sub> and represent the averages of three replicates; error bars, standard deviations. ]] |
Revision as of 10:18, 7 November 2010
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UNIQ207d3be797f45a18-partinfo-00000000-QINU
No review score entered. iGEM Tokyo_Tech 2010 |
In order to characterize R0061, Plux repression promoter, we constructed K395101 combining R0061 and K121013, which is a promoter-less gfp reporter (rbs-gfp-ter-ter) on pSB6A1 and used a fusion of PlacIq (I14032) to gfp (K121013) as a positive control and used promoterless gfp (K121013) as a negative control. |
iGEM Chiba 2010 |
iGEM Chiba 2010
DetailsMaterials & Methods
Results
Reference<biblio>
</biblio> |
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