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TUDelft/16 September 2008
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==Colony PCR 3A Assembly== | ==Colony PCR 3A Assembly== | ||
| - | The transformation of the overnight ligation of the available parts K115029-K115036 didn't work on most plates. Only K115032 and K115034 had 1 and 3 colonies respectively. K115036 might have had a colony but was not very convincing. Colony PCR has been done on these spots, showing only K115034 was at the right size. New restrictions and ligations have been started for all parts K115029-K115036 (excluding K115030 and K115035 as not all DNA has arrived yet), again we will use vector pSB1AT3. | + | The transformation of the overnight ligation of the available parts K115029-K115036 didn't work on most plates. Only K115032 and K115034 had 1 and 3 colonies respectively. K115036 might have had a colony but was not very convincing. Colony PCR has been done on these spots, showing only K115034 was at the right size. New restrictions and ligations have been started for all parts K115029-K115036 (excluding K115030 and K115035 as not all DNA has arrived yet), again we will use vector pSB1AT3. The blank showed a curious result, but later results showed our water is clean. We did stir a toothpick through the colony PCR mix blank sample, which might have been infected somehow. |
| - | ==Gel of the E. coli genes== | + | [[Image:080916.png|thumb|250px|center|Figure 1. Gel containing the final construct of K115034, indicated by the arrow.]] |
| + | |||
| + | ==Gel of the ''E. coli'' genes== | ||
It seems most of the transformants were closed vectors without insert. Currently, we could test more colonies and hope there are some good ones in there. On the other hand, it may be worthwhile and a more certain way to go to order new primers (containing the whole prefix and suffix, allowing for ''Pst''I & ''Eco''RI cutting) to avoid the assembly problem we are now facing. XbaI and SpeI digestions yield compatible sticky ends... | It seems most of the transformants were closed vectors without insert. Currently, we could test more colonies and hope there are some good ones in there. On the other hand, it may be worthwhile and a more certain way to go to order new primers (containing the whole prefix and suffix, allowing for ''Pst''I & ''Eco''RI cutting) to avoid the assembly problem we are now facing. XbaI and SpeI digestions yield compatible sticky ends... | ||
{{Template:TUDelftiGEM2008_sidebar}} | {{Template:TUDelftiGEM2008_sidebar}} | ||
Latest revision as of 16:29, 28 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 16th
Colony PCR 3A Assembly
The transformation of the overnight ligation of the available parts K115029-K115036 didn't work on most plates. Only K115032 and K115034 had 1 and 3 colonies respectively. K115036 might have had a colony but was not very convincing. Colony PCR has been done on these spots, showing only K115034 was at the right size. New restrictions and ligations have been started for all parts K115029-K115036 (excluding K115030 and K115035 as not all DNA has arrived yet), again we will use vector pSB1AT3. The blank showed a curious result, but later results showed our water is clean. We did stir a toothpick through the colony PCR mix blank sample, which might have been infected somehow.
Gel of the E. coli genes
It seems most of the transformants were closed vectors without insert. Currently, we could test more colonies and hope there are some good ones in there. On the other hand, it may be worthwhile and a more certain way to go to order new primers (containing the whole prefix and suffix, allowing for PstI & EcoRI cutting) to avoid the assembly problem we are now facing. XbaI and SpeI digestions yield compatible sticky ends...
