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Team:Newcastle/Labwork/18 September 2009
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# 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], i.e. ''cotC-GFP-smtA'', ''kinA'', ''pGFP-rrnB'', ''pMUTIN4'' and ''pSB1AT3'' | # 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], i.e. ''cotC-GFP-smtA'', ''kinA'', ''pGFP-rrnB'', ''pMUTIN4'' and ''pSB1AT3'' | ||
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===Procedure=== | ===Procedure=== | ||
| + | ====Starting ''Bacillus subtilis'' transformations==== | ||
| + | ====Designing primers for ''cotC'' BioBrick integration==== | ||
| + | ====Plating out TPA2 cells==== | ||
| + | ====Analysing midi-prep digsts==== | ||
| + | ====Analysing ''cotC'', ''kinA'', ''pGFP-rrnB'', ''pMUTIN4'' and ''pSB1AT3''==== | ||
| + | <br> | ||
<center> | <center> | ||
{|cellpadding="10" | {|cellpadding="10" | ||
Revision as of 15:45, 13 October 2009
Project
Sub-projects
Wet Lab
Meetings
Planning
Links
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, i.e. cotC-GFP-smtA, kinA, pGFP-rrnB, pMUTIN4 and pSB1AT3
Procedure
Starting Bacillus subtilis transformations
Designing primers for cotC BioBrick integration
Plating out TPA2 cells
Analysing midi-prep digsts
Analysing cotC, kinA, pGFP-rrnB, pMUTIN4 and pSB1AT3
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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]
