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TUDelft/11 September 2008
From 2008.igem.org
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{{Template:TUDelftiGEM2008_calendar}} | {{Template:TUDelftiGEM2008_calendar}} | ||
| - | =September 11th | + | =September 11th= |
==Transformation of PCR products== | ==Transformation of PCR products== | ||
| - | Ligation of PCR products of E. coli genes was performed o/n and a direct transformation of the ligation products was performed. 50 ul of competent cells were added to the 15 ul ligation mix transformed o/n. Results will be checked by colony PCR tomorrow. | + | Ligation of PCR products of ''E. coli'' genes was performed o/n and a direct transformation of the ligation products was performed. 50 ul of competent cells were added to the 15 ul ligation mix transformed o/n. Results will be checked by colony PCR tomorrow. |
==Colony PCR of 3A Assembly== | ==Colony PCR of 3A Assembly== | ||
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The transformants from the assembly have grown, indicating the assembly has worked. We did a colony PCR on a couple of the transformants, to check for insert sizes. The gel of the colony PCR is displayed in figure 1. | The transformants from the assembly have grown, indicating the assembly has worked. We did a colony PCR on a couple of the transformants, to check for insert sizes. The gel of the colony PCR is displayed in figure 1. | ||
[[Image:TUDelft110908cPCR.jpg|thumb|center|Figure 1. Colony PCR of the thermosensitive part / luciferase+Terminator 3A assembly, the lanes indicated by arrows were used to continue assembly, or in the case of K115012, to use as a negative control for the experiments.]] | [[Image:TUDelft110908cPCR.jpg|thumb|center|Figure 1. Colony PCR of the thermosensitive part / luciferase+Terminator 3A assembly, the lanes indicated by arrows were used to continue assembly, or in the case of K115012, to use as a negative control for the experiments.]] | ||
| - | We suspect the parts which | + | We suspect the parts which gave multiple bands at very different sizes are parts which have not ligated inserts but are just a couple of backbones ligated together, as this would give primer annealing zones in all kinds of orientations. For each assembly we've PCR'd a construct of the correct size (some extra bands could be a [http://partsregistry.org/Problems_with_PCR_using_VR/VF2 consequence] of the use of B0015 as terminator). |
{{Template:TUDelftiGEM2008_sidebar}} | {{Template:TUDelftiGEM2008_sidebar}} | ||
Latest revision as of 10:41, 27 October 2008
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| [http://2008.igem.org/TUDelft/1_September_2008 1] | [http://2008.igem.org/TUDelft/2_September_2008 2] | [http://2008.igem.org/TUDelft/3_September_2008 3] | [http://2008.igem.org/TUDelft/4_September_2008 4] | [http://2008.igem.org/TUDelft/5_September_2008 5] | [http://2008.igem.org/wiki/index.php?title=TUDelft/6_September_2008&action=edit 6] | [http://2008.igem.org/wiki/index.php?title=TUDelft/7_September_2008&action=edit 7] |
| [http://2008.igem.org/TUDelft/8_September_2008 8] | [http://2008.igem.org/TUDelft/9_September_2008 9] | [http://2008.igem.org/TUDelft/10_September_2008 10] | [http://2008.igem.org/TUDelft/11_September_2008 11] | [http://2008.igem.org/TUDelft/12_September_2008 12] | [http://2008.igem.org/wiki/index.php?title=TUDelft/13_September_2008&action=edit 13] | [http://2008.igem.org/wiki/index.php?title=TUDelft/14_September_2008&action=edit 14] |
| [http://2008.igem.org/TUDelft/15_September_2008 15] | [http://2008.igem.org/TUDelft/16_September_2008 16] | [http://2008.igem.org/TUDelft/17_September_2008 17] | [http://2008.igem.org/TUDelft/18_September_2008 18] | [http://2008.igem.org/TUDelft/19_September_2008 19] | [http://2008.igem.org/wiki/index.php?title=TUDelft/20_September_2008&action=edit 20] | [http://2008.igem.org/wiki/index.php?title=TUDelft/21_September_2008&action=edit 21] |
| [http://2008.igem.org/TUDelft/22_September_2008 22] | [http://2008.igem.org/TUDelft/23_September_2008 23] | [http://2008.igem.org/TUDelft/24_September_2008 24] | [http://2008.igem.org/TUDelft/25_September_2008 25] | [http://2008.igem.org/wiki/index.php?title=TUDelft/26_September_2008&action=edit 26] | [http://2008.igem.org/wiki/index.php?title=TUDelft/27_September_2008&action=edit 27] | [http://2008.igem.org/wiki/index.php?title=TUDelft/28_September_2008&action=edit 28] |
| [http://2008.igem.org/TUDelft/29_September_2008 29] | [http://2008.igem.org/TUDelft/30_September_2008 30] | |||||
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| [http://2008.igem.org/TUDelft/1_October_2008 1] | [http://2008.igem.org/TUDelft/2_October_2008 2] | [http://2008.igem.org/TUDelft/3_October_2008 3] | [http://2008.igem.org/wiki/index.php?title=TUDelft/4_October_2008&action=edit 4] | [http://2008.igem.org/wiki/index.php?title=TUDelft/5_October_2008&action=edit 5] | ||
| [http://2008.igem.org/TUDelft/6_October_2008 6] | [http://2008.igem.org/TUDelft/7_October_2008 7] | [http://2008.igem.org/TUDelft/8_October_2008 8] | [http://2008.igem.org/TUDelft/9_October_2008 9] | [http://2008.igem.org/TUDelft/10_October_2008 10] | [http://2008.igem.org/wiki/index.php?title=TUDelft/11_October_2008&action=edit 11] | [http://2008.igem.org/wiki/index.php?title=TUDelft/12_October_2008&action=edit 12] |
| [http://2008.igem.org/TUDelft/13_October_2008 13] | [http://2008.igem.org/TUDelft/14_October_2008 14] | [http://2008.igem.org/TUDelft/15_October_2008 15] | [http://2008.igem.org/TUDelft/16_October_2008 16] | [http://2008.igem.org/TUDelft/17_October_2008 17] | [http://2008.igem.org/wiki/index.php?title=TUDelft/18_October_2008&action=edit 18] | [http://2008.igem.org/wiki/index.php?title=TUDelft/19_October_2008&action=edit 19] |
| [http://2008.igem.org/TUDelft/20_October_2008 20] | [http://2008.igem.org/TUDelft/21_October_2008 21] | [http://2008.igem.org/TUDelft/22_October_2008 22] | [http://2008.igem.org/TUDelft/23_October_2008 23] | [http://2008.igem.org/TUDelft/24_October_2008 24] | [http://2008.igem.org/wiki/index.php?title=TUDelft/25_October_2008&action=edit 25] | [http://2008.igem.org/wiki/index.php?title=TUDelft/26_October_2008&action=edit 26] |
| [http://2008.igem.org/wiki/index.php?title=TUDelft/27_October_2008&action=edit 27] | [http://2008.igem.org/wiki/index.php?title=TUDelft/28_October_2008&action=edit 28] | [http://2008.igem.org/wiki/index.php?title=TUDelft/29_October_2008&action=edit 29] | [http://2008.igem.org/wiki/index.php?title=TUDelft/30_October_2008&action=edit 30] | [http://2008.igem.org/wiki/index.php?title=TUDelft/31_October_2008&action=edit 31] | ||
September 11th
Transformation of PCR products
Ligation of PCR products of E. coli genes was performed o/n and a direct transformation of the ligation products was performed. 50 ul of competent cells were added to the 15 ul ligation mix transformed o/n. Results will be checked by colony PCR tomorrow.
Colony PCR of 3A Assembly
The transformants from the assembly have grown, indicating the assembly has worked. We did a colony PCR on a couple of the transformants, to check for insert sizes. The gel of the colony PCR is displayed in figure 1.
We suspect the parts which gave multiple bands at very different sizes are parts which have not ligated inserts but are just a couple of backbones ligated together, as this would give primer annealing zones in all kinds of orientations. For each assembly we've PCR'd a construct of the correct size (some extra bands could be a [http://partsregistry.org/Problems_with_PCR_using_VR/VF2 consequence] of the use of B0015 as terminator).
