Team:Uppsala-SwedenWeek14

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(→Construction Of K1, K2, K3)
 
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== Construction Of K1, K2, K3 ==
== Construction Of K1, K2, K3 ==
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The samples inoculated previously were taken out and glecerol stocks were made.
The inoclum obtained from the overnight culture was used for plasmid extraction.  
The inoclum obtained from the overnight culture was used for plasmid extraction.  
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The extracted plasmids were cut at specific restriction sites present in the biobrick sequence and run on the gel.
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The extracted plasmids were cut at specific restriction sites present in the biobrick sequence and run on the gel.`
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The gel was checked for correct fragment lengths to perform a second round f validation
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The gel images of the bio-bricks with the band lengths were shown below in
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The extracted plasmids were digested with the correct set of enzymes as defined in the flow chart.
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Fig1, FIg2, Fig3
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The extracted plasmids were purified and used for performing ligation with the correct prefix or suffix and the appropriate vector.
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[[Image:20100906 K1 and K2 cPCR 20100902 (2).jpg|500px|center|border|thumb|K1-K2 Figure 1]]
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The ligate was used to perform transformation of competent cells
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The competent cells were plated on appropriate cells for transformation.
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The concentration of plasmid were measured for all the samples.
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[[Image:20100906 K1 and K2 cPCR 20100902 thumb.jpg|500px|center|border|thumb|K1-K2 Figure 2]]
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Plasmid concentration of all starting biobricks.Plasmid of J1, J2, J3 were digested with proper enzymes. The biobricks were ligated in pairs according to the plan by Quick Ligase kit from Fermentas.
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[[Image:20100906 K3 cPCR 20100902 (2) thumb.jpg|500px|center|border|thumb|K3 Figure 3]]
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The ligation product were transformed into DH5α competent cells and each sample were plated on the agar plate which antibiotics matched to its backbone antibiotic resistance.
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The gel was checked for correct fragment lengths to perform a second round f validation
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The extracted plasmids were digested with the correct set of enzymes as defined in the flow chart.
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The concentration of plasmid were measured for all the samples.

Latest revision as of 21:12, 26 October 2010

Week-14

Construction Of K1, K2, K3

The samples inoculated previously were taken out and glecerol stocks were made.

The inoclum obtained from the overnight culture was used for plasmid extraction. The extracted plasmids were cut at specific restriction sites present in the biobrick sequence and run on the gel.`

The gel images of the bio-bricks with the band lengths were shown below in Fig1, FIg2, Fig3

K1-K2 Figure 1
K1-K2 Figure 2
K3 Figure 3


The gel was checked for correct fragment lengths to perform a second round f validation The extracted plasmids were digested with the correct set of enzymes as defined in the flow chart.

The concentration of plasmid were measured for all the samples.

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