Team:UNIPV-Pavia/Notebook/Week5Jul
From 2009.igem.org
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(→July, 27th) |
(→July, 27th) |
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*Gel results: | *Gel results: | ||
- | **Samples 1, | + | **Samples 1, 4, 10, 11, 14, 19 and 20 showed an extra band for the non ligated plasmid; |
- | **Samples 2, | + | **Samples 2, 9, 13 and 18 were pure! we decided to keep B1-13 (lane 8) to perform future ligations. |
- | *NOTE: we had a pure sample for B1 and | + | *NOTE: we had a pure sample for B1 (i.e. B1-13)and three almost-pure sample for B2. Anyway, we decided to perform ligation reactions for these samples and the extra band of B2 will be eliminated during gel cut/purification. WE DECIDED TO KEEP B2-5. Next weeks we will think about purifying B2-5 itself. |
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+ | |||
+ | *We transformed 20 pg of B1-13 purified DNA (stored at -20°C) in TOP10 in order to prepare a glycerol stock for this construct. We incubated the plate at 37°C overnight. | ||
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''Preparation of experiment with Tecan F200'' | ''Preparation of experiment with Tecan F200'' | ||
- | *We infected 5 ml of LB + Amp with 10 ul of A14pg | + | *We infected 5 ml of LB + Amp with 10 ul of A14pg, A8pg and A9pg glycerol stocks. |
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+ | *We also infected 5 ml of LB + Amp with a single colony taken from B0030 native plate (stored at +4°C). | ||
- | *We incubated the | + | *We incubated the inocula at 37°C, 220 rpm overnight. |
Revision as of 13:03, 4 August 2009
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Week from July 27th, to August 1st, 2009
Previous Week | Next Week |
July, 27th
- Screening for the 11 miniprepped DNA samples for B1: digestion S-P for all.
- Gel results:
- Samples 1, 4, 10, 11, 14, 19 and 20 showed an extra band for the non ligated plasmid;
- Samples 2, 9, 13 and 18 were pure! we decided to keep B1-13 (lane 8) to perform future ligations.
- NOTE: we had a pure sample for B1 (i.e. B1-13)and three almost-pure sample for B2. Anyway, we decided to perform ligation reactions for these samples and the extra band of B2 will be eliminated during gel cut/purification. WE DECIDED TO KEEP B2-5. Next weeks we will think about purifying B2-5 itself.
- We transformed 20 pg of B1-13 purified DNA (stored at -20°C) in TOP10 in order to prepare a glycerol stock for this construct. We incubated the plate at 37°C overnight.
Preparation of experiment with Tecan F200
- We infected 5 ml of LB + Amp with 10 ul of A14pg, A8pg and A9pg glycerol stocks.
- We also infected 5 ml of LB + Amp with a single colony taken from B0030 native plate (stored at +4°C).
- We incubated the inocula at 37°C, 220 rpm overnight.
July, 28th
July, 29th
July, 30th
July, 31st
Previous Week | Next Week |