Difference between revisions of "Team:BIOSINT Mexico/Project"
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<h3>spisPink & amilGFP optimized to Escherichia coli K12 </h3> | <h3>spisPink & amilGFP optimized to Escherichia coli K12 </h3> | ||
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<br>After completing the insertion of gBlocks (IDT) designed (spisPink and amilGFP), into the plasmid pSB1C3, by following the relevant protocols; competent cells were successfully transformed and consequently the removal of plasmids (spisPink into pSB1C3) BBa_K1684000 and (amilGFP into pSB1C3) BBa_K1684001 was performed.</br> | <br>After completing the insertion of gBlocks (IDT) designed (spisPink and amilGFP), into the plasmid pSB1C3, by following the relevant protocols; competent cells were successfully transformed and consequently the removal of plasmids (spisPink into pSB1C3) BBa_K1684000 and (amilGFP into pSB1C3) BBa_K1684001 was performed.</br> | ||
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<br>The Figure 6 presents a 1% agarose gel which contains two samples, by triplicate, next to the ladder. In line 1, 2 and 3 are located the triplicate samples of spisPink piece, optimized for E. coli K12 (BBa_K1684000), with a size of 2789 bp. Likewise on the line 4,5 and 6 are repeated samples of the piece amilGFP optimized for E. coli K12 (BBa_K1684001). For both BioBricks, the electrophoresis results correspond to the theoretical size, greater than 2500 bp and less than 3000 bp; also it can be seen that there is no variation between samples, replicas, indicating the reliability of E. coli K12 transformations.</br> | <br>The Figure 6 presents a 1% agarose gel which contains two samples, by triplicate, next to the ladder. In line 1, 2 and 3 are located the triplicate samples of spisPink piece, optimized for E. coli K12 (BBa_K1684000), with a size of 2789 bp. Likewise on the line 4,5 and 6 are repeated samples of the piece amilGFP optimized for E. coli K12 (BBa_K1684001). For both BioBricks, the electrophoresis results correspond to the theoretical size, greater than 2500 bp and less than 3000 bp; also it can be seen that there is no variation between samples, replicas, indicating the reliability of E. coli K12 transformations.</br> | ||
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<h3>RBS and Optimized biobricks</h3> | <h3>RBS and Optimized biobricks</h3> | ||
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<br>In Figure 7 different samples of parts used for transformation of competent cells; these two parts are composite biobricks, the first is composed by RBS (BBa_B0034) and the optimized chromoprotein sequence spisPink for E. coli K12 (BBa_K1684000), while the second is composed by a RBS (BBa_B0034) and the optimized chromoprotein sequence amilGFP (BBa_K1684001 ). </br> | <br>In Figure 7 different samples of parts used for transformation of competent cells; these two parts are composite biobricks, the first is composed by RBS (BBa_B0034) and the optimized chromoprotein sequence spisPink for E. coli K12 (BBa_K1684000), while the second is composed by a RBS (BBa_B0034) and the optimized chromoprotein sequence amilGFP (BBa_K1684001 ). </br> | ||
<br>The agarose gel shown in Line 1, the negative control; Line 2, belongs to the purified DNA of pGLO (5371 bp), was run as a positive control. Line 3 and Line 4 contain duplicate products of BBa_K1684002 (BBa_B0034 + BBa_K1684000) biobrick, both results match the size of the plasmids, 2800 bp. Line 5 and Line 6 contain duplicate BBa_K1684003 (BBa_B0034 + BBa_K1684001) biobrick; however, a positive result is observed only in Line 5, with size of 2822 bp, while in line 6 only presents contamination, without the presence of any genetic material. </br> | <br>The agarose gel shown in Line 1, the negative control; Line 2, belongs to the purified DNA of pGLO (5371 bp), was run as a positive control. Line 3 and Line 4 contain duplicate products of BBa_K1684002 (BBa_B0034 + BBa_K1684000) biobrick, both results match the size of the plasmids, 2800 bp. Line 5 and Line 6 contain duplicate BBa_K1684003 (BBa_B0034 + BBa_K1684001) biobrick; however, a positive result is observed only in Line 5, with size of 2822 bp, while in line 6 only presents contamination, without the presence of any genetic material. </br> | ||
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<h3>Optimized biobricks with Terminator</h3> | <h3>Optimized biobricks with Terminator</h3> | ||
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<br>Finally, in Figure 8 samples composite parts are presented, the first BBa_K1684004, is composed of the chromoprotein sequence optimized for E. coli K12 spisPink (BBa_K1684000) and a terminator (BBa_B0010); the second part is composed of the chromoprotein optimized sequence amilGFP (BBa_K1684001) and the same terminator (BBa_B0010). This agarose gel shown on Line 1 and Line 2, the biobrick BBa_K1684004, in the first case the band run corresponding to its size, 2869 bp; however, on Line 2 the result was not the expected and only contamination was obtained.</br> | <br>Finally, in Figure 8 samples composite parts are presented, the first BBa_K1684004, is composed of the chromoprotein sequence optimized for E. coli K12 spisPink (BBa_K1684000) and a terminator (BBa_B0010); the second part is composed of the chromoprotein optimized sequence amilGFP (BBa_K1684001) and the same terminator (BBa_B0010). This agarose gel shown on Line 1 and Line 2, the biobrick BBa_K1684004, in the first case the band run corresponding to its size, 2869 bp; however, on Line 2 the result was not the expected and only contamination was obtained.</br> | ||
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<br>In Line 3 and 4 results, was run the composite BioBrick BBa_K1684005, in the first case the result was not successful; In Line 4 however, the band is present with the respective size, 2890 bp, as a positive result. Finally on Line 5 is presented only the negative control.</br> | <br>In Line 3 and 4 results, was run the composite BioBrick BBa_K1684005, in the first case the result was not successful; In Line 4 however, the band is present with the respective size, 2890 bp, as a positive result. Finally on Line 5 is presented only the negative control.</br> | ||
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Revision as of 07:31, 18 September 2015
Coliroid
Background
Overview
Color Coliroid
Assembly
Methods
Light Cannon
Results
spisPink & amilGFP optimized to Escherichia coli K12
After completing the insertion of gBlocks (IDT) designed (spisPink and amilGFP), into the plasmid pSB1C3, by following the relevant protocols; competent cells were successfully transformed and consequently the removal of plasmids (spisPink into pSB1C3) BBa_K1684000 and (amilGFP into pSB1C3) BBa_K1684001 was performed.
The Figure 6 presents a 1% agarose gel which contains two samples, by triplicate, next to the ladder. In line 1, 2 and 3 are located the triplicate samples of spisPink piece, optimized for E. coli K12 (BBa_K1684000), with a size of 2789 bp. Likewise on the line 4,5 and 6 are repeated samples of the piece amilGFP optimized for E. coli K12 (BBa_K1684001). For both BioBricks, the electrophoresis results correspond to the theoretical size, greater than 2500 bp and less than 3000 bp; also it can be seen that there is no variation between samples, replicas, indicating the reliability of E. coli K12 transformations.
RBS and Optimized biobricks
In Figure 7 different samples of parts used for transformation of competent cells; these two parts are composite biobricks, the first is composed by RBS (BBa_B0034) and the optimized chromoprotein sequence spisPink for E. coli K12 (BBa_K1684000), while the second is composed by a RBS (BBa_B0034) and the optimized chromoprotein sequence amilGFP (BBa_K1684001 ).
The agarose gel shown in Line 1, the negative control; Line 2, belongs to the purified DNA of pGLO (5371 bp), was run as a positive control. Line 3 and Line 4 contain duplicate products of BBa_K1684002 (BBa_B0034 + BBa_K1684000) biobrick, both results match the size of the plasmids, 2800 bp. Line 5 and Line 6 contain duplicate BBa_K1684003 (BBa_B0034 + BBa_K1684001) biobrick; however, a positive result is observed only in Line 5, with size of 2822 bp, while in line 6 only presents contamination, without the presence of any genetic material.
Optimized biobricks with Terminator
Finally, in Figure 8 samples composite parts are presented, the first BBa_K1684004, is composed of the chromoprotein sequence optimized for E. coli K12 spisPink (BBa_K1684000) and a terminator (BBa_B0010); the second part is composed of the chromoprotein optimized sequence amilGFP (BBa_K1684001) and the same terminator (BBa_B0010). This agarose gel shown on Line 1 and Line 2, the biobrick BBa_K1684004, in the first case the band run corresponding to its size, 2869 bp; however, on Line 2 the result was not the expected and only contamination was obtained.
In Line 3 and 4 results, was run the composite BioBrick BBa_K1684005, in the first case the result was not successful; In Line 4 however, the band is present with the respective size, 2890 bp, as a positive result. Finally on Line 5 is presented only the negative control.
After completing the insertion of gBlocks (IDT) designed (spisPink and amilGFP), into the plasmid pSB1C3, by following the relevant protocols; competent cells were successfully transformed and consequently the removal of plasmids (spisPink into pSB1C3) BBa_K1684000 and (amilGFP into pSB1C3) BBa_K1684001 was performed.
The Figure 6 presents a 1% agarose gel which contains two samples, by triplicate, next to the ladder. In line 1, 2 and 3 are located the triplicate samples of spisPink piece, optimized for E. coli K12 (BBa_K1684000), with a size of 2789 bp. Likewise on the line 4,5 and 6 are repeated samples of the piece amilGFP optimized for E. coli K12 (BBa_K1684001). For both BioBricks, the electrophoresis results correspond to the theoretical size, greater than 2500 bp and less than 3000 bp; also it can be seen that there is no variation between samples, replicas, indicating the reliability of E. coli K12 transformations.
In Figure 7 different samples of parts used for transformation of competent cells; these two parts are composite biobricks, the first is composed by RBS (BBa_B0034) and the optimized chromoprotein sequence spisPink for E. coli K12 (BBa_K1684000), while the second is composed by a RBS (BBa_B0034) and the optimized chromoprotein sequence amilGFP (BBa_K1684001 ).
The agarose gel shown in Line 1, the negative control; Line 2, belongs to the purified DNA of pGLO (5371 bp), was run as a positive control. Line 3 and Line 4 contain duplicate products of BBa_K1684002 (BBa_B0034 + BBa_K1684000) biobrick, both results match the size of the plasmids, 2800 bp. Line 5 and Line 6 contain duplicate BBa_K1684003 (BBa_B0034 + BBa_K1684001) biobrick; however, a positive result is observed only in Line 5, with size of 2822 bp, while in line 6 only presents contamination, without the presence of any genetic material.
Finally, in Figure 8 samples composite parts are presented, the first BBa_K1684004, is composed of the chromoprotein sequence optimized for E. coli K12 spisPink (BBa_K1684000) and a terminator (BBa_B0010); the second part is composed of the chromoprotein optimized sequence amilGFP (BBa_K1684001) and the same terminator (BBa_B0010). This agarose gel shown on Line 1 and Line 2, the biobrick BBa_K1684004, in the first case the band run corresponding to its size, 2869 bp; however, on Line 2 the result was not the expected and only contamination was obtained.
In Line 3 and 4 results, was run the composite BioBrick BBa_K1684005, in the first case the result was not successful; In Line 4 however, the band is present with the respective size, 2890 bp, as a positive result. Finally on Line 5 is presented only the negative control.