Difference between revisions of "Part:BBa K1412829:Experience"

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[[File:Verification.png |820px|thumb|center|<b>Figure 1</b> Verification of plasmid BBa_K1412614 and BBa_K1412014.]]
  
 
==='''Activiation'''===
 
==='''Activiation'''===
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1. Transfer 50 μL bacterium solution (pLac-RBS (1.0)-cheZ-TT, pLac-RBS (0.01)-cheZ-TT, pLac-RBS (0.3)-cheZ-TT) into 5 ml new LB liquid medium whose chloromycetin concentration is 50ug/ml to culture for 12 hours in 37℃ horizontal rotators at 200 rpm.
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1. Transfer 50 μL bacterium solution (pLac-RBS(1.0)-<i>cheZ</i>-TT, pLac-RBS(0.01)-<i>cheZ</i>-TT, pLac-RBS(0.3)-<i>cheZ</i>-TT) into 5 ml new LB liquid medium whose chloromycetin concentration is 50ug/ml to culture for 12 hours in 37℃ horizontal rotators at 200 rpm.
  
 
2. Then transfer another 50μL bacterium solution from the LB above to another new 5ml LB liquid medium whose chloromycetin concentration is 50ug/ml to culture in 37℃ horizontal rotators at 200rpm for 3 hours.
 
2. Then transfer another 50μL bacterium solution from the LB above to another new 5ml LB liquid medium whose chloromycetin concentration is 50ug/ml to culture in 37℃ horizontal rotators at 200rpm for 3 hours.
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3. Finally, culture the bacteria in constant temperature and humidity incubator at 37℃ and observe the condition of bacterium growth.
 
3. Finally, culture the bacteria in constant temperature and humidity incubator at 37℃ and observe the condition of bacterium growth.
  
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[[File:Xmu_project_application_RBSpromoter02.png |810px|thumb|center|<b>Figure 2</b>. The schematic diagram of how we measure the diameter of colonies.]]
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[[File:Schematic_diagram.jpg |750px|thumb|center|<b>Figure 2</b>. The schematic diagram of how we measure the diameter of colonies.]]
  
 
===='''Measurement'''====
 
===='''Measurement'''====
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R2, R3, R4, R5, R6, R7….
 
R2, R3, R4, R5, R6, R7….
  
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===Applications of BBa_K1412801===
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===Applications of BBa_K1412829===
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We use this part to characterize the efficiency of different ribosome binding sites.
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===User Reviews===
 
===User Reviews===
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Latest revision as of 00:34, 18 October 2014


Protocol

Verification

Figure 1 Verification of plasmid BBa_K1412614 and BBa_K1412014.

Activiation

1. Transfer 50 μL bacterium solution (pLac-RBS(1.0)-cheZ-TT, pLac-RBS(0.01)-cheZ-TT, pLac-RBS(0.3)-cheZ-TT) into 5 ml new LB liquid medium whose chloromycetin concentration is 50ug/ml to culture for 12 hours in 37℃ horizontal rotators at 200 rpm.

2. Then transfer another 50μL bacterium solution from the LB above to another new 5ml LB liquid medium whose chloromycetin concentration is 50ug/ml to culture in 37℃ horizontal rotators at 200rpm for 3 hours.

Culture & Measurement

Culture

1. Firstly, draw three dots on a plate before using oily pen from the bottom of culture dish.

2. Secondly, stab 3μl bacterium medium into the M63 semisolid medium at the dots.

3. Finally, culture the bacteria in constant temperature and humidity incubator at 37℃ and observe the condition of bacterium growth.

Figure 2. The schematic diagram of how we measure the diameter of colonies.

Measurement

1. Observe the the condition of bacterium growth,and prepare a ruler.

2. When we can see a visible difference of the diameter of semi-solid medium with time pass by, begin to measure the diameter of bacterium colonies from the bottom of the semi-solid medium to avoiding the collapse of semi-solid medium.

3. Recording data: The diameter of initial colonies(0h) is recorded as R1. And the diameter of the colonies at 12h, 24h, 30h, 36h, 42h... as R2, R3, R4, R5, R6, R7….

Applications of BBa_K1412801

User Reviews

UNIQ61cb69833200be58-partinfo-00000000-QINU UNIQ61cb69833200be58-partinfo-00000001-QINU