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Edinburgh/30 July 2008
From 2008.igem.org
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(New page: <html> <head> <title>Edinburgh iGEM 2008</title> <script type="text/javascript"> //Drop Down Tabs Menu- Author: Dynamic Drive (http://www.dynamicdrive.com) //Created: May 16th, 07' var ta...) |
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| - | * '''P48''' (''crtY'') and '''P49''' ( | + | </style> |
| - | * Ligated | + | </html> |
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| + | {| align="left" | ||
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| + | |align="left" width="150pt"|{{#calendar: title=Edinburgh |year=2008 | month=06}} | ||
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| + | |align="left" width="150pt"|{{#calendar: title=Edinburgh |year=2008 | month=07}} | ||
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| + | |align="left" width="150pt"|{{#calendar: title=Edinburgh |year=2008 | month=08}} | ||
| + | |} | ||
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| + | :::: '''[[Edinburgh/29_July_2008|< Previous Entry]]''' | ||
| + | |||
| + | == Week 7 == | ||
| + | === Wednesday 30 July 08 === | ||
| + | |||
| + | * '''P48''' (''crtY'' using a new primer mixture) and '''P49''' (recreation of P12, rbs+''dxs'') created. (CF) | ||
| + | * Ligated putative ''cenA'' and putative ''cex'' fragments to Edinbrick 1('''L28''' and '''L29''' respectively). All were digested with EcoRI/SpeI, purified and stored overnight in the 16°C water bath. (AM) | ||
* Repeated PCR for ''cenA'' and ''cex'' ('''P46''', '''P47''') from heat killed cell solution. Denaturing 95°C for 1 min, annealing 65°C for 10s, extension 70°C for 40s. Run on '''Gel 30'''. Unsuccessful, perhaps because annealing temperature was too high (temperature was decided based on stock solution label, which took the prefix/suffix into consideration). (AM) | * Repeated PCR for ''cenA'' and ''cex'' ('''P46''', '''P47''') from heat killed cell solution. Denaturing 95°C for 1 min, annealing 65°C for 10s, extension 70°C for 40s. Run on '''Gel 30'''. Unsuccessful, perhaps because annealing temperature was too high (temperature was decided based on stock solution label, which took the prefix/suffix into consideration). (AM) | ||
| - | * | + | * Digested M49 and M50 (pSB1A2-rbs+''crtI'') with a) XbaI, b) SpeI/XbaI, c) sac/speI, M63 (BABEL2+rbs+''crtE'') with a) EcoRI, b) EcoRI/PstI, M67 (pSB1A2+rbs+''crtB'') with a) EcoRI/PstI, b) sacI/speI and ran on '''Gel 31''' with P48 (''crtY''). (OG) |
| - | * Re-digested M55 | + | * Re-digested M55-M60 (pSB1A2+pZntA) with a) EcoRI and b) EcoRI/PstI. (This time, the amount of DNA for digestion is increased) and ran on '''gel 32'''. Results look promising for M57, but not for the others. (Yan) |
| - | + | * Purified P15 (rbs+''appY'') and P28 (''crtB''), digested each with Edinbrick1 using XbaI/PstI and set-up for ligations ('''L26''' and '''L27'''). Stored overnight in the 16°C water bath. (HX)<br /> | |
| - | * Purified P15 (rbs+''appY'') and P28 (''crtB''), digested each with | + | <br /> |
| + | |||
| + | :::: '''[[Edinburgh/31_July_2008|Next Entry >]]''' | ||
Revision as of 15:33, 26 August 2008
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Week 7
Wednesday 30 July 08
- P48 (crtY using a new primer mixture) and P49 (recreation of P12, rbs+dxs) created. (CF)
- Ligated putative cenA and putative cex fragments to Edinbrick 1(L28 and L29 respectively). All were digested with EcoRI/SpeI, purified and stored overnight in the 16°C water bath. (AM)
- Repeated PCR for cenA and cex (P46, P47) from heat killed cell solution. Denaturing 95°C for 1 min, annealing 65°C for 10s, extension 70°C for 40s. Run on Gel 30. Unsuccessful, perhaps because annealing temperature was too high (temperature was decided based on stock solution label, which took the prefix/suffix into consideration). (AM)
- Digested M49 and M50 (pSB1A2-rbs+crtI) with a) XbaI, b) SpeI/XbaI, c) sac/speI, M63 (BABEL2+rbs+crtE) with a) EcoRI, b) EcoRI/PstI, M67 (pSB1A2+rbs+crtB) with a) EcoRI/PstI, b) sacI/speI and ran on Gel 31 with P48 (crtY). (OG)
- Re-digested M55-M60 (pSB1A2+pZntA) with a) EcoRI and b) EcoRI/PstI. (This time, the amount of DNA for digestion is increased) and ran on gel 32. Results look promising for M57, but not for the others. (Yan)
- Purified P15 (rbs+appY) and P28 (crtB), digested each with Edinbrick1 using XbaI/PstI and set-up for ligations (L26 and L27). Stored overnight in the 16°C water bath. (HX)
