"
Team:Newcastle/Labwork/27 July 2009
From 2009.igem.org
(Difference between revisions)
(→Preparation of the plates) |
|||
| (27 intermediate revisions not shown) | |||
| Line 2: | Line 2: | ||
{{:Team:Newcastle/Header}} | {{:Team:Newcastle/Header}} | ||
{{:Team:Newcastle/Left}} | {{:Team:Newcastle/Left}} | ||
| + | __NOTOC__ | ||
| + | [[Image:Team Newcastle 2009 iGEM ProbationaryP-Sign.PNG|50px|right]] | ||
| + | =Formal Lab Session - 27th July 2009= | ||
| + | [[Image:Team Newcastle 2009 iGEM 27-07-09 IMG 0178.JPG|250px|center]] | ||
| - | |||
| - | Today we prepared the gel for last week's rescued plasmids which contain RFP and GFP biobricks | + | == Running the gel for RFP and GFP bioricks== |
| + | Today we prepared the gel with 0.8% agorose for last week's rescued plasmids which contain RFP and GFP biobricks. The final solution was 500ml using 4 gr of agorose in total. | ||
| - | We checked the agar plates containing colonies of plasmids with five different biobricks from last week. | + | We mixed 9ul of DNas with 1ul of the buffer and centrifuged shortly. The rest of the DNAs are then put back to the freezer. |
| - | The list of plates and the biobricks are below: | + | |
| + | Finally we run the gel. We loaded the ladder to the first and fourth wells, GFP to the 2nd and RFP to the 3rd well and set the voltage to 120V which was then reduced to 90V. | ||
| + | |||
| + | Below are the images: | ||
| + | [[Image:Team Newcastle - iGem2009 - geldoc 2009-07-27.jpg|center]] | ||
| + | |||
| + | |||
| + | ==Transformation of new biobricks== | ||
| + | We prepared the agar plates and left the tubes including the biobricks for incubation overnight. | ||
| + | |||
| + | We checked the agar plates containing colonies of plasmids with five different biobricks from last week. We will be following [https://2009.igem.org/Team:Newcastle/Project/Labwork/PhilsProtocols#Phil.E2.80.99s_mini_method_for_Alkaline_Lysis_for_Mini_Prep Phil’s mini method for Alkaline Lysis for Mini Prep] The list of plates and the biobricks are below: | ||
{| class="wikitable" border="1" | {| class="wikitable" border="1" | ||
| Line 14: | Line 28: | ||
| Plate | | Plate | ||
| BioBrick | | BioBrick | ||
| - | | Number | + | | Number used for the labelling |
| + | | Name | ||
|- | |- | ||
| Plate 1 | | Plate 1 | ||
| - | | | + | | [http://partsregistry.org/Part:BBa_C0056 BBa_C0056] |
| 1 | | 1 | ||
| + | | CI Coding Sequence | ||
|- | |- | ||
| Plate 2 | | Plate 2 | ||
| - | | | + | | [http://partsregistry.org/Part:BBa_B1002 BBa_B1002] |
| 2 | | 2 | ||
| + | | Terminator | ||
|- | |- | ||
| Plate 3 | | Plate 3 | ||
| - | | | + | | [http://partsregistry.org/Part:BBa_C0077 BBa_C0077] |
| 3 | | 3 | ||
| + | | CinR Coding sequence | ||
|- | |- | ||
| Plate 4 | | Plate 4 | ||
| - | | | + | | [http://partsregistry.org/Part:BBa_C0076 BBa_C0076] |
| 4 | | 4 | ||
| + | | CinI Coding Sequence | ||
|- | |- | ||
| Plate 5 | | Plate 5 | ||
| - | | | + | | [http://partsregistry.org/Part:BBa_R0077 BBa_R0077] |
| 5 | | 5 | ||
| + | | CinR sensitive promoter | ||
|- | |- | ||
|} | |} | ||
| - | + | Except from plate 3, we had colonies formed in all plates. | |
Colonies from the plates were picked and mixed in a tube containing 5ml of LB. We prepared 12 tubes for 4 different biobricks. For each biobrick we labelled the tubes as A, B, and C. Finally we left them at shaking incubator overnight. | Colonies from the plates were picked and mixed in a tube containing 5ml of LB. We prepared 12 tubes for 4 different biobricks. For each biobrick we labelled the tubes as A, B, and C. Finally we left them at shaking incubator overnight. | ||
==Preparation of the plates== | ==Preparation of the plates== | ||
| - | We prepared the agar solutions | + | [[Image:Team Newcastle 2009 iGEM 27-07-09 IMG 0177.JPG|200px|thumb|Goksel pouring out agar plates]] |
| + | We prepared the agar solutions and poured into the plates which are then placed into the fridge. | ||
The list of plates and their content are as below. | The list of plates and their content are as below. | ||
| Line 49: | Line 70: | ||
|- | |- | ||
| Content | | Content | ||
| - | | Number of plates | + | | Number of plates prepared |
| + | | Labelled as | ||
|- | |- | ||
| LB | | LB | ||
| 4 | | 4 | ||
| + | | LB 27/07/09 | ||
|- | |- | ||
| LB + Amp | | LB + Amp | ||
| 6 | | 6 | ||
| + | | LB+A 27/07/09 | ||
|- | |- | ||
| LB + Amp + Kan | | LB + Amp + Kan | ||
| 5 | | 5 | ||
| + | | LB+AK 27/07/09 | ||
|- | |- | ||
|} | |} | ||
| + | |||
| + | |||
| + | ===Tomorrow=== | ||
| + | We have placed the required solutions to the bench for the experiment apart from RNAase. We need to find it first thing in the morning tomorrow then we will follow [https://2009.igem.org/Team:Newcastle/Project/Labwork/PhilsProtocols#Phil.E2.80.99s_mini_method_for_Alkaline_Lysis_for_Mini_Prep Phil’s mini method for Alkaline Lysis for Mini Prep]. GFP and RFP are placed into the freezer. The DNA ladder is in the fridge. Plates are placed into the fridge just below the top shelf. They were split into two since they did not all fit to the shelf. | ||
| + | |||
| + | {{:Team:Newcastle/Project/Labwork/CalTemplate}} | ||
| + | |||
{{:Team:Newcastle/Footer}} | {{:Team:Newcastle/Footer}} | ||
{{:Team:Newcastle/Right}} | {{:Team:Newcastle/Right}} | ||
Latest revision as of 17:21, 17 October 2009
Project
Sub-projects
Wet Lab
Meetings
Planning
Links
Formal Lab Session - 27th July 2009
Running the gel for RFP and GFP bioricks
Today we prepared the gel with 0.8% agorose for last week's rescued plasmids which contain RFP and GFP biobricks. The final solution was 500ml using 4 gr of agorose in total.
We mixed 9ul of DNas with 1ul of the buffer and centrifuged shortly. The rest of the DNAs are then put back to the freezer.
Finally we run the gel. We loaded the ladder to the first and fourth wells, GFP to the 2nd and RFP to the 3rd well and set the voltage to 120V which was then reduced to 90V.
Below are the images:
Transformation of new biobricks
We prepared the agar plates and left the tubes including the biobricks for incubation overnight.
We checked the agar plates containing colonies of plasmids with five different biobricks from last week. We will be following Phil’s mini method for Alkaline Lysis for Mini Prep The list of plates and the biobricks are below:
| Plate | BioBrick | Number used for the labelling | Name |
| Plate 1 | [http://partsregistry.org/Part:BBa_C0056 BBa_C0056] | 1 | CI Coding Sequence |
| Plate 2 | [http://partsregistry.org/Part:BBa_B1002 BBa_B1002] | 2 | Terminator |
| Plate 3 | [http://partsregistry.org/Part:BBa_C0077 BBa_C0077] | 3 | CinR Coding sequence |
| Plate 4 | [http://partsregistry.org/Part:BBa_C0076 BBa_C0076] | 4 | CinI Coding Sequence |
| Plate 5 | [http://partsregistry.org/Part:BBa_R0077 BBa_R0077] | 5 | CinR sensitive promoter |
Except from plate 3, we had colonies formed in all plates.
Colonies from the plates were picked and mixed in a tube containing 5ml of LB. We prepared 12 tubes for 4 different biobricks. For each biobrick we labelled the tubes as A, B, and C. Finally we left them at shaking incubator overnight.
Preparation of the plates
We prepared the agar solutions and poured into the plates which are then placed into the fridge. The list of plates and their content are as below.
| Content | Number of plates prepared | Labelled as |
| LB | 4 | LB 27/07/09 |
| LB + Amp | 6 | LB+A 27/07/09 |
| LB + Amp + Kan | 5 | LB+AK 27/07/09 |
Tomorrow
We have placed the required solutions to the bench for the experiment apart from RNAase. We need to find it first thing in the morning tomorrow then we will follow Phil’s mini method for Alkaline Lysis for Mini Prep. GFP and RFP are placed into the freezer. The DNA ladder is in the fridge. Plates are placed into the fridge just below the top shelf. They were split into two since they did not all fit to the shelf.
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
News
Events
- 20 – 21 June 2009 - Europe workshop (London)
- 23 – 24 June 2009 - UK iGEM meetup (Edinburgh)
- 23 October Practice Presentation (Newcastle)
- 23 October T-shirts are ready
- 27 October Practice Presentation (Sunderland)
- 27 October Poster is ready
- 30 October – 2 November 2009 - Jamboree (Boston)
Social Net
- Newcastle iGEM Twitter
- [http://www.facebook.com/home.php#/group.php?gid=131709337641 Newcastle on Facebook]
- [http://www.youtube.com/user/newcastle2009igem Newcastle Youtube Channel]
