Team:Newcastle/Labwork/18 September 2009
From 2009.igem.org
(Difference between revisions)
(→Practical Outline) |
(→Practical Outline) |
||
Line 22: | Line 22: | ||
<br> | <br> | ||
# Start the ''Bacillus subtilis'' transformation process ([https://2009.igem.org/Team:Newcastle/Project/Labwork/OurProtocols/TransformBac#On_the_day_of_Bacillus_subtilis_transformations_.28starting_in_the_morning.29 "Day of transformation" steps] of protocol) in the morning. | # Start the ''Bacillus subtilis'' transformation process ([https://2009.igem.org/Team:Newcastle/Project/Labwork/OurProtocols/TransformBac#On_the_day_of_Bacillus_subtilis_transformations_.28starting_in_the_morning.29 "Day of transformation" steps] of protocol) in the morning. | ||
+ | # Design primers for checking ''B. subtilis'' ''cotC'' integration | ||
+ | # Plate out some TPA2 cells for production of competent ''Dh5-alpha E. coli'' cells | ||
+ | # Run digests from yesterday (midi-prep digests) on gel | ||
+ | ## If successful transform ''B. subtilis'' with midi-prep DNA | ||
+ | ## If unsuccessful prepare another ligation attempt | ||
+ | # Carry out a second attempt at digesting the 5 midi-prep samples from the [https://2009.igem.org/Team:Newcastle/Labwork/11_September_2009#Midi-prep_cotC-GFP-smtA.2C_kinA.2C_pGFP-rrnB.2C_pMUTIN4_and_pSB1AT3 11/09/09 Lab Session]. | ||
===Procedure=== | ===Procedure=== |
Revision as of 12:57, 13 October 2009
Formal Lab Session - 18th September 2009
Stochastic Switch Team
Today we did midipreps of the ara transformation and digested 10ul to run on a gel (EcoRI + PstI) Two bands could be seen one for the backbone and one correctly sied for the insert (~200bp) so this midiprep can be taken forward in order to do the ara-sspb double clone.
Today we also made up some fresh DNA ladder as we have had problems with degradation.
- 40ul of 500ug/ml promega lambda HindIII digest (20ug in total)
- 280ul H2O
- 80ul loading dye
=400ul which was aliquoted into 4 tubes; 3 put in the freezer 1 in the fridge.
Metal Sensing Team
Introduction
Practical Outline
This is the initial list of tasks which need to be carried out by the end of the day:
- Start the Bacillus subtilis transformation process ("Day of transformation" steps of protocol) in the morning.
- Design primers for checking B. subtilis cotC integration
- Plate out some TPA2 cells for production of competent Dh5-alpha E. coli cells
- Run digests from yesterday (midi-prep digests) on gel
- If successful transform B. subtilis with midi-prep DNA
- If unsuccessful prepare another ligation attempt
- Carry out a second attempt at digesting the 5 midi-prep samples from the 11/09/09 Lab Session.
Procedure
|
|
|
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]