Team:UCL/Labbook/Week10

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Lab Weeks

Week 10

Bacterial Lab

5th August - The new batch of ampicillin was tested via transformation of competent cells with pSecTag2A. Plates were spread and incubated 37C o/n.

6th August - Results from yesterday’s 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.

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 amp powder was located and amp was remade.

8th August - Preparation of 4x Amp plates and 4x no drug plates for storage in the fridge.

9th August - Meeting with Darren Nesbeth, requirements for upcoming weeks: create glycerol stocks of both pSecTag2A and pSB1C3, purify and extract pure DNA for pSecTag2A and pSB1C3.

Mammalian Lab

6th August - Checked HeLa cells. Cells are growing slow, left to grow for a few more days

8th August - HeLa cells are about 30% confluent. Changed media.