Team:Paris Liliane Bettencourt/Notebook/2010/09/05/
From 2010.igem.org
Aleksandra
Touchdown PCR of the 13 minipreps from yesterday and also of 6 other constructs (elongation time 6 mins), then gel electrophoresis.
Gel 1 (PCR)
Minipreps from 31/08
- 1 Pms/wTet II
- 2 Pms/mTet III
- 3 Psm/mTet III
- 4 Psw/mTet III
- 5 Pss/wTet II
- 6 Psw/wTet II
Minipreps from yesterday (04/05)
- 7,8 Pms/mTet/LuxI, 2 cultures
- 9,10 Pss/wTet/LuxI, 2 cultures
- 11,12 Psm/sTet/LuxI, 2 cultures
- 13,14 Psm/wTetIII/I13507/pLux/GFP, 2 cultures
- 15,16 Psm/wTetII/I13507/pLux/GFP, 2 cultures
- Ladder 1 kb
<a href="/wiki/Image:PCR0509.jpg" class="image" title="Image:PCR0509.jpg"><img alt="Image:PCR0509.jpg" src="/images/thumb/4/49/PCR0509.jpg/687px-PCR0509.jpg.png" width="687" height="386" border="0" /></a>
Gel 2 (PCR)
Minipreps from yesterday (04/05)
- 17, 18 Pss/sLac/LuxI III, 2 cultures
- 19 Pms/sTet/LuxI
Digestion from today(see further)
- 19 Pms/sTet/LuxI
<a href="/wiki/Image:PCR0509_2.jpg" class="image" title="Image:PCR0509_2.jpg"><img alt="Image:PCR0509_2.jpg" src="/images/thumb/0/0c/PCR0509_2.jpg/335px-PCR0509_2.jpg.png" width="335" height="400" border="0" /></a>
→ Colonies 7, 9, 10, 17, 18, 19 seem to be of a right size
Restriction digest and gel electrophoresis of yesterday's minipreps with E+P
Gel 1
- 1 attP/PmsRFP, col 1
- Ladder 1kb
- 2 attP/PmsRFP, col 2
- -
- 3 attP/PsmRFP, col 1
- -
- 4 attP/PsmRFP, col 2
- -
- 5 attP/PssRFP, col 1
- -
- 6 attP/PssRFP, col 2
- -
- 7 Pms/mTet/LuxI, col 1
- -
- 8 Pms/mTet/LuxI, col 2
- -
- 9 Pss/wTet/LuxI, col 1
<a href="/wiki/Image:Gel0509_2uncut.jpg" class="image" title="Image:gel0509_2uncut.jpg"><img alt="Image:gel0509_2uncut.jpg" src="/images/thumb/0/0d/Gel0509_2uncut.jpg/682px-Gel0509_2uncut.jpg.png" width="682" height="408" border="0" /></a>
Gel 2
- 10 Pss/wTet/LuxI, col 2
- -
- 11 Psm/sTet/LuxI, col 1
- -
- 12 Psm/sTet/LuxI, col 2
- -
- 13 Psm/wTetIII/I13507/pLux/GFP, col 1
- -
- 14 Psm/wTetIII/I13507/pLux/GFP, col 1
- -
- 15 Psm/wTetII/I13507/pLux/GFP, col 1
- -
- 16 Psm/wTetII/I13507/pLux/GFP, col 2
- -
- 17 Pss/sLac/LuxI III, col 1
- -
- 18 Pss/sLac/LuxI III, col 2
<a href="/wiki/Image:Gel0509_1uncut.jpg" class="image" title="Image:gel0509_1uncut.jpg"><img alt="Image:gel0509_1uncut.jpg" src="/images/thumb/5/5b/Gel0509_1uncut.jpg/684px-Gel0509_1uncut.jpg.png" width="684" height="399" border="0" /></a>
<a href="/wiki/Image:Gel0509_1cut.jpg" class="image" title="Image:gel0509_1cut.jpg"><img alt="Image:gel0509_1cut.jpg" src="/images/thumb/0/01/Gel0509_1cut.jpg/690px-Gel0509_1cut.jpg.png" width="690" height="388" border="0" /></a>
- 19 Pms/sTet/LuxI (on the PCR gel, see above)
→ Digestions 1, 2, 7, 9, 10, 17, 18, 19 seem to be of a right size