Team:UNIPV-Pavia/Notebook/Week3Oct
From 2009.igem.org
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== <html><font class="dayw_style">October, 12th</font></html> == | == <html><font class="dayw_style">October, 12th</font></html> == | ||
+ | *We repeated the digestion for F2620TOP10(S-P) and B5new2-3(X-P-ClaI). | ||
+ | *Gel run/cut for the two plasmids: bands were good. | ||
+ | |||
+ | *Gel extraction for: | ||
+ | **F2620TOP10(S-P) | ||
+ | **B5new2-3(X-P-ClaI) | ||
+ | **B3(X-P) | ||
+ | **B4(X-P) | ||
+ | **A19-1(S-P) | ||
+ | |||
+ | *Unfortunately we had a very low DNA yield for almost all of them: A19-1(S-P) and F2620(S-P) could not be used :'( | ||
+ | |||
+ | *We decided not to repeat A19-1 extraction (and then the A21 re-cloning) and to assay our A21 construct for ethanol production. | ||
+ | |||
+ | *F2620 DNA was over, so we inoculated 8 ul of F2620MIT1 glycerol stock in 7 ml of LB + Amp. We incubated this inoculum overnight (37°C, 220 rpm). | ||
+ | |||
+ | *We inoculated 12 of our assemblies in 3 ml of LB + Amp in order to prepare the DNA to send to iGEM HQ: | ||
+ | **A1 | ||
+ | **A2 | ||
+ | **A3 | ||
+ | **A4 | ||
+ | **A5 | ||
+ | **A6 | ||
+ | **A7 | ||
+ | **A8pg | ||
+ | **A9pg | ||
+ | **A11-3 | ||
+ | **A12-2 | ||
+ | **A15-3 | ||
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Revision as of 09:33, 15 October 2009
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Week from October 12th, to October 18th, 2009
Previous Week | Next Week |
October, 12th
- We repeated the digestion for F2620TOP10(S-P) and B5new2-3(X-P-ClaI).
- Gel run/cut for the two plasmids: bands were good.
- Gel extraction for:
- F2620TOP10(S-P)
- B5new2-3(X-P-ClaI)
- B3(X-P)
- B4(X-P)
- A19-1(S-P)
- Unfortunately we had a very low DNA yield for almost all of them: A19-1(S-P) and F2620(S-P) could not be used :'(
- We decided not to repeat A19-1 extraction (and then the A21 re-cloning) and to assay our A21 construct for ethanol production.
- F2620 DNA was over, so we inoculated 8 ul of F2620MIT1 glycerol stock in 7 ml of LB + Amp. We incubated this inoculum overnight (37°C, 220 rpm).
- We inoculated 12 of our assemblies in 3 ml of LB + Amp in order to prepare the DNA to send to iGEM HQ:
- A1
- A2
- A3
- A4
- A5
- A6
- A7
- A8pg
- A9pg
- A11-3
- A12-2
- A15-3
October, 13th
- Miniprep for the 12 inocula to be sent.
- Miniprep, cut S-P, gel run/cut/purification for F2620MIT1. This time we had a good yield;)
- Ligations:
- B14 = F2620(S-P) + B3(X-P) in pSB1A2
- B15 = F2620(S-P) + B4(X-P) in pSB1A2
- We incubated them at 16°C overnight.
October, 14th
- We transformed the two ligations (after 1:20 dilution) and incubated the plated bacteria overnight (37°C).
- We inoculated all the remaining assemblies (12) in order to send purified DNA to iGEM HQ (3 ml of LB + Amp).
- We inoculated 8 ul of B5new2-3, F2620TOP10 and B0015 glycerol stocks in 8 ml of LB + Amp + 2% glucose. We incubated them at 37°C, 220 rpm in the morning.
- Team meeting
October, 15th
October, 16th
October, 17th
October, 18th
Previous Week | Next Week |