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Team:Uppsala-SwedenWeek13
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{{Template:Uppsala}}<!--Do not remove the first and last lines in this page!--> | {{Template:Uppsala}}<!--Do not remove the first and last lines in this page!--> | ||
== Week-13 == | == Week-13 == | ||
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Identification of biobrciks | Identification of biobrciks | ||
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Plasmid of the samples which showed right size band on the gel were sent to a plasmid concentration test. We decided only J1, J2, J3 were in good concentration to proceed. | Plasmid of the samples which showed right size band on the gel were sent to a plasmid concentration test. We decided only J1, J2, J3 were in good concentration to proceed. | ||
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However, we got the correct PCR lengths for Bio-bricks and results are shown in image files below. | However, we got the correct PCR lengths for Bio-bricks and results are shown in image files below. | ||
| - | .The result was showed in Fig.1 | + | .The result was showed in Fig.1 and Fig2 |
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| - | [[Image:20100827 J1-J3 c-PCR 20100826 2 thumb.jpg| | + | J1-J3 |
| + | [[Image:20100827 J1-J3 c-PCR 20100826 2 thumb.jpg|500px|center|border|Figure 1]] | ||
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| + | J2-J3 | ||
| + | [[Image:20100827 J2-J3 c-PCR 20100826 2 thumb.jpg|500px|center|border|Figure 1]] | ||
biobricks | biobricks | ||
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However, we got the correct band lengths for all J1, J2, J3 shows the band with right size on the gel pic. | However, we got the correct band lengths for all J1, J2, J3 shows the band with right size on the gel pic. | ||
J1, J2, J3 samples were again inoculated again to get the enough plasmid for further progress. | J1, J2, J3 samples were again inoculated again to get the enough plasmid for further progress. | ||
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Second colony PCR was performed again for all the starting biobricks and the products were run for gel again. Comparing the DNA length to the band size, all the biobricks were confirmed. | Second colony PCR was performed again for all the starting biobricks and the products were run for gel again. Comparing the DNA length to the band size, all the biobricks were confirmed. | ||
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Table 2 Plasmid concentration of all starting biobricks. | Table 2 Plasmid concentration of all starting biobricks. | ||
Plasmid of J1, J2, J3 were digested with proper enzymes bought from Sigma-Aldrich. The biobricks were ligated in pairs according to the plan by Quick Ligase kit from Fermentas. | Plasmid of J1, J2, J3 were digested with proper enzymes bought from Sigma-Aldrich. The biobricks were ligated in pairs according to the plan by Quick Ligase kit from Fermentas. | ||
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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. | 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. | ||
Revision as of 19:37, 26 October 2010
Week-13
Identification of biobrciks
Plasmid of the samples which showed right size band on the gel were sent to a plasmid concentration test. We decided only J1, J2, J3 were in good concentration to proceed.
However, we got the correct PCR lengths for Bio-bricks and results are shown in image files below. .The result was showed in Fig.1 and Fig2
J1-J3
J2-J3
biobricks
However, we got the correct band lengths for all J1, J2, J3 shows the band with right size on the gel pic. J1, J2, J3 samples were again inoculated again to get the enough plasmid for further progress.
Second colony PCR was performed again for all the starting biobricks and the products were run for gel again. Comparing the DNA length to the band size, all the biobricks were confirmed.
The concentration of plasmid were measured for all the samples and values were showed in Table 2 Table 2 Plasmid concentration of all starting biobricks. Plasmid of J1, J2, J3 were digested with proper enzymes bought from Sigma-Aldrich. The biobricks were ligated in pairs according to the plan by Quick Ligase kit from Fermentas.
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.
