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Thursday, August 18
From 2011.igem.org
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Tomorrow at 1:00pm I will check this page, and the lab will open at the earliest time that '''at least 2''' iGEMers intend to arrive. An email to the googlegroup will be sent, and the opening time, although it should be obvious to all, will be posted at the top of this page. | Tomorrow at 1:00pm I will check this page, and the lab will open at the earliest time that '''at least 2''' iGEMers intend to arrive. An email to the googlegroup will be sent, and the opening time, although it should be obvious to all, will be posted at the top of this page. | ||
| - | |||
| - | + | 1. On 8/4/11 we performed colony PCR on the two colonies (one from 3B and one from 4A) that we obtained by transformation on 7/31/11. Today we are running a gel of the PCR products that were amplified by colony PCR on 8/4/11. We prepared 1 % gel and did the colony PCR from 3B and 4A. | |
| - | We | + | |
| - | + | Gel Electrophoresis of PCR Reaction 8.4.11 | |
| + | We used 6 lanes and ran 25 ul of PCR products (except for the bench top ladder): | ||
| + | *Lanes 1 and 6 are 6 ul of Bench top ladder.(600 ul 100lanes, 1kb) | ||
| - | 2 -ve control | + | *Lane 2 -ve control (is this positive control?) |
| - | 3 3B | + | *Lane 3 3B |
| - | + | *Lane 4 4A | |
| - | 5 No template | + | *Lane 5 No template |
The gel was started at 9.14 at 82 volt and stopped at 10.10. | The gel was started at 9.14 at 82 volt and stopped at 10.10. | ||
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| + | |||
| + | 2. We checked the results of the transformation from 8/4/11. Results: | ||
| + | *Negative control: no colonies | ||
| + | *3B undiluted: 9 colonies | ||
| + | *3B diluted 1:10: 2 colonies | ||
| + | *4A (plate says from 50 ul of transformation): >100 colonies | ||
| + | *4A undiluted: 3 colonies | ||
| + | |||
| + | We set up colony PCR from these colonies in case none of the colonies from the 8/4/11 colony PCR are correct. After these PCR reactions are completed, they need to be run on a gel and then, if there is product, cut with PstI. We ran 24 PCR reaction (a negative control with no template DNA, a positive control using the pET-Taq plasmid as a template, and 22 reactions using bacteria from the plates as templates). | ||
| + | |||
| + | |||
| + | 3. We also checked the results of the transformation of Biobrick parts from the registry from 8/4/11. We had no colonies for either the part from the 5I or 5K wells. Dr. Burkett will work on repeating the transformation in his lab in case there was a problem with the cells or the antibiotics in the plates. | ||
Revision as of 16:51, 22 August 2011
The Lab Opens Today at 5:00 pm. Could someone please email our googlegroup to this effect? For some reason I am unable to sign into my GMail or TU accounts!. Dr Liz
The time the Lab will this Thursday will depend upon the responses to this request: Please add your name and the time you intend to come to the lab on this day. Put an asterisk(*) before your line to add it to the list. Tomorrow at 1:00pm I will check this page, and the lab will open at the earliest time that at least 2 iGEMers intend to arrive. An email to the googlegroup will be sent, and the opening time, although it should be obvious to all, will be posted at the top of this page.
1. On 8/4/11 we performed colony PCR on the two colonies (one from 3B and one from 4A) that we obtained by transformation on 7/31/11. Today we are running a gel of the PCR products that were amplified by colony PCR on 8/4/11. We prepared 1 % gel and did the colony PCR from 3B and 4A.
Gel Electrophoresis of PCR Reaction 8.4.11 We used 6 lanes and ran 25 ul of PCR products (except for the bench top ladder):
- Lanes 1 and 6 are 6 ul of Bench top ladder.(600 ul 100lanes, 1kb)
- Lane 2 -ve control (is this positive control?)
- Lane 3 3B
- Lane 4 4A
- Lane 5 No template
The gel was started at 9.14 at 82 volt and stopped at 10.10.
Rocking started at 10.10 at 50 rpm.
2. We checked the results of the transformation from 8/4/11. Results:
- Negative control: no colonies
- 3B undiluted: 9 colonies
- 3B diluted 1:10: 2 colonies
- 4A (plate says from 50 ul of transformation): >100 colonies
- 4A undiluted: 3 colonies
We set up colony PCR from these colonies in case none of the colonies from the 8/4/11 colony PCR are correct. After these PCR reactions are completed, they need to be run on a gel and then, if there is product, cut with PstI. We ran 24 PCR reaction (a negative control with no template DNA, a positive control using the pET-Taq plasmid as a template, and 22 reactions using bacteria from the plates as templates).
3. We also checked the results of the transformation of Biobrick parts from the registry from 8/4/11. We had no colonies for either the part from the 5I or 5K wells. Dr. Burkett will work on repeating the transformation in his lab in case there was a problem with the cells or the antibiotics in the plates.
Person/ Time
- Dr Liz Time = to be determined
- Travis; approximately 5pm
- Steve: soonest today would be 7:00 PM
- Bivor: From 12 and to 10.30pm
- Desirae: 2-9pm
