Team:UCL/Labbook/Week10

From 2013.igem.org

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<b>Monday 5th August</b> - The new batch of <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> ampicillin</a> was tested via <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> transformation</a> of <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> competent cells</a> with pSecTag2A. Plates were <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> spread</a> and incubated 37C o/n.
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<b>Monday 5th August</b>
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<b>Tuesday 6th August</b> - Results from yesterday’s transformation:
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The new batch of <a href="https://2013.igem.org/Team:UCL/Project/Protocols"> ampicillin</a> was tested via <a href="https://2013.igem.org/Team:UCL/Project/Protocols"> transformation</a> of competent cells with pSecTag2A. Plates were <a href="https://2013.igem.org/Team:UCL/Project/Protocols"> spread</a> and incubated at 37°C overnight.
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<b>Tuesday 6th August</b>
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Results from 5th August transformation:
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This indicates that there is still an issue with the <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> ampicillin</a>, possibly a problem with the stock powder used. A final test with both old and new <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> ampicillin</a> was carried out and compared without plasmid insertion.  
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This indicates that there is still an issue with the <a href="https://2013.igem.org/Team:UCL/Project/Protocols"> ampicillin</a>, possibly a problem with the stock powder used. A final test with both old and new <a href="https://2013.igem.org/Team:UCL/Project/Protocols"> ampicillin</a> was carried out and compared without plasmid insertion.  
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<b>Wednesday 7th August</b> -
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<b>Wednesday 7th August</b>  
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Results indicated that the <a href="https://2013.igem.org/Team:UCL/Project/Protocols"> ampicillin</a> source may not have been fully functional, therefore a new source of ampicillin powder was located and ampicillin was remade.
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Results indicated that the <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> ampicillin</a> source may not have been fully functional, therefore a new source of amp powder was located and amp was remade.
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<b>Thursday 8th August</b>
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<b>Thursday 8th August</b> - Preparation of 4x <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> Amp plates</a> and 4x <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> no drug plates</a> for storage in the fridge.
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Preparation of 4x <a href="https://2013.igem.org/Team:UCL/Project/Protocols"> ampicillin plates</a> and 4x <a href="https://2013.igem.org/Team:UCL/Project/Protocols"> no drug plates</a> for storage in the fridge.
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<b>Friday 9th August</b> - Meeting with Darren Nesbeth, requirements for upcoming weeks: create <a href="https://2013.igem.org/Team:UCL/Project/Protocols" target="_blank"> glycerol stocks</a> of both pSecTag2A and pSB1C3, purify and extract pure DNA for pSecTag2A and pSB1C3.
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<b>Friday 9th August</b>
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Meeting with supervisor Dr Darren Nesbeth
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Goals to complete for upcoming weeks:
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create <a href="https://2013.igem.org/Team:UCL/Project/Protocols"> glycerol stocks</a> of both pSecTag2A and pSB1C3, purify and extract pure DNA for pSecTag2A and pSB1C3.
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<b>Tuesday 6th August</b> - Checked HeLa cells. Cells are growing slow, left to grow for a few more days
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<b>Tuesday 6th August</b>
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Checked HeLa cells. Cells are growing slow, left to grow for a few more days
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<b>Thursday 8th August</b>
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<b>Thursday 8th August</b> - HeLa cells are about 30% confluent. Changed media.
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HeLa cells are about 30% confluent. Changed media.
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Latest revision as of 19:13, 4 October 2013

Lab Weeks

Week 10

Bacterial Lab

Monday 5th August

The new batch of ampicillin was tested via transformation of competent cells with pSecTag2A. Plates were spread and incubated at 37°C overnight.

Tuesday 6th August

Results from 5th August transformation:

Vial Ampicillin Plate Plasmid Insertion Colony Count
1 (Main) Yes Yes 100+
2 (Positive Control) No Yes 100+
3 (Negative Control) Yes No 15

This indicates that there is still an issue with the ampicillin, possibly a problem with the stock powder used. A final test with both old and new ampicillin was carried out and compared without plasmid insertion.

Wednesday 7th August

Vial Ampicillin Plate Plasmid Insertion Colony Count
1 Old Ampicillin Yes Yes 100+
2 New Ampicillin v2 No Yes 100+
3 Positive Control Yes No 15

Results indicated that the ampicillin source may not have been fully functional, therefore a new source of ampicillin powder was located and ampicillin was remade.

Thursday 8th August

Preparation of 4x ampicillin plates and 4x no drug plates for storage in the fridge.

Friday 9th August

Meeting with supervisor Dr Darren Nesbeth

Goals to complete for upcoming weeks:

create glycerol stocks of both pSecTag2A and pSB1C3, purify and extract pure DNA for pSecTag2A and pSB1C3.

Mammalian Lab

Tuesday 6th August

Checked HeLa cells. Cells are growing slow, left to grow for a few more days

Thursday 8th August

HeLa cells are about 30% confluent. Changed media.