Difference between revisions of "Part:BBa K1412614:Experience"
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====Verification==== | ====Verification==== | ||
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1. Transfer pBAD-RBS-<i>CheZ</i>-TT (<bbpart>BBa_K1412624</bbpart>) into competent cell of <i>E.coli</i> (ΔCheZ) respectively. | 1. Transfer pBAD-RBS-<i>CheZ</i>-TT (<bbpart>BBa_K1412624</bbpart>) into competent cell of <i>E.coli</i> (ΔCheZ) respectively. | ||
| − | 2. Coated plates(LB solid medium with the antibiotic concentration 50ug/ml chloromycetin) and culture in 37℃ biochemical incubator for 12h. | + | 2. Coated plates(LB solid medium with the antibiotic concentration 50ug/ml chloromycetin) and culture in 37℃ biochemical incubator for |
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| + | 12h. | ||
3. Select colony to culture in 5ml LB fluid medium, of which the chloromycetin, antibiotic concentration is 50ug/ml. | 3. Select colony to culture in 5ml LB fluid medium, of which the chloromycetin, antibiotic concentration is 50ug/ml. | ||
| − | 4. Activation of bacterium: transfer 50uL bacterium medium into a new 5ml LB fluid medium, of which the chloromycetin, antibiotic concentration is 50ug/ml. | + | 4. Activation of bacterium: transfer 50uL bacterium medium into a new 5ml LB fluid medium, of which the chloromycetin, antibiotic |
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| + | concentration is 50ug/ml. | ||
5. Stab of bacterium: We draw three spots on a plate before, then stab 3ul bacterium medium into the M63 semisolid medium at the spots. | 5. Stab of bacterium: We draw three spots on a plate before, then stab 3ul bacterium medium into the M63 semisolid medium at the spots. | ||
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====Measurement==== | ====Measurement==== | ||
| − | 1. The initial colony radius is recorded as R1, and the radius are measured at 12h, 24h, 30h, 36h, 42h... as R2, R3, R4, R5, R6, … The migration radius caused by chemotaxis grows higher over time, larger radius bringing less error. Each radius minus R1 is the net migration radius at certain time. Plot the net chemotaxis radius versus time and set the radius of pLac-RBS (1.0)-CheZ-TT (<bbpart>BBa_K1412000</bbpart>) as RBS strength 1.0. Then we can calculate the RBS efficiency (ΔR2L/ΔR2M) and (R2J/ΔR2M) and compare them with the official data. | + | 1. The initial colony radius is recorded as R1, and the radius are measured at 12h, 24h, 30h, 36h, 42h... as R2, R3, R4, R5, R6, … The |
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| + | migration radius caused by chemotaxis grows higher over time, larger radius bringing less error. Each radius minus R1 is the net migration | ||
| + | |||
| + | radius at certain time. Plot the net chemotaxis radius versus time and set the radius of pLac-RBS (1.0)-CheZ-TT | ||
| + | |||
| + | (<bbpart>BBa_K1412000</bbpart>) as RBS strength 1.0. Then we can calculate the RBS efficiency (ΔR2L/ΔR2M) and (R2J/ΔR2M) and compare | ||
| + | |||
| + | them with the official data. | ||
===Applications of BBa_K1412614=== | ===Applications of BBa_K1412614=== | ||
Revision as of 14:30, 16 October 2014
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Protocol
Verification
Characterization stage
1. Transfer pBAD-RBS-CheZ-TT (BBa_K1412624) into competent cell of E.coli (ΔCheZ) respectively.
2. Coated plates(LB solid medium with the antibiotic concentration 50ug/ml chloromycetin) and culture in 37℃ biochemical incubator for
12h.
3. Select colony to culture in 5ml LB fluid medium, of which the chloromycetin, antibiotic concentration is 50ug/ml.
4. Activation of bacterium: transfer 50uL bacterium medium into a new 5ml LB fluid medium, of which the chloromycetin, antibiotic
concentration is 50ug/ml.
5. Stab of bacterium: We draw three spots on a plate before, then stab 3ul bacterium medium into the M63 semisolid medium at the spots.
6. Culture the bacteria in constant temperature and humidity incubator at 37℃.
Measurement
1. The initial colony radius is recorded as R1, and the radius are measured at 12h, 24h, 30h, 36h, 42h... as R2, R3, R4, R5, R6, … The
migration radius caused by chemotaxis grows higher over time, larger radius bringing less error. Each radius minus R1 is the net migration
radius at certain time. Plot the net chemotaxis radius versus time and set the radius of pLac-RBS (1.0)-CheZ-TT
(BBa_K1412000) as RBS strength 1.0. Then we can calculate the RBS efficiency (ΔR2L/ΔR2M) and (R2J/ΔR2M) and compare
them with the official data.
Applications of BBa_K1412614
User Reviews
UNIQf3678669ccad6118-partinfo-00000002-QINU UNIQf3678669ccad6118-partinfo-00000003-QINU