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Edinburgh/29 July 2008
From 2008.igem.org
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* Cut out putative ''cenA'' and ''cex'' (both ~1.5kb) fragments from '''gel 27''' (repeat of Gel 26 but stained with SYBRsafe). Purified DNA from gel fragments. (AM) | * Cut out putative ''cenA'' and ''cex'' (both ~1.5kb) fragments from '''gel 27''' (repeat of Gel 26 but stained with SYBRsafe). Purified DNA from gel fragments. (AM) | ||
* Digests of M49-M54 (pSB1A2+rbs+''crtI'') with EcoRI alone. This should generate a single 4 kb band (doing an EcoRI/PstI double digest would give a vector and insert band both around 2 kb, which would probably run at the same place, making it hard to tell whether there was an insert). These were run on '''Gel 28'''. All showed a single band at the expected place. (CF) | * Digests of M49-M54 (pSB1A2+rbs+''crtI'') with EcoRI alone. This should generate a single 4 kb band (doing an EcoRI/PstI double digest would give a vector and insert band both around 2 kb, which would probably run at the same place, making it hard to tell whether there was an insert). These were run on '''Gel 28'''. All showed a single band at the expected place. (CF) | ||
| - | * Minipreps of pSB1A2+ | + | * Minipreps of pSB1A2+''P<sub>zntA</sub>'' ('''M55-M60''') from L23, pSB1A2+rbs+''crtE'' ('''M61-M66''') from L22 and pSB1A2+rbs+''crtB'' ('''M67''') from L21. (Yan, Nimisha) |
| - | ** Double digests of M55-M66 with EcoRI/PstI. This should generate the following bands: ~2.2 kb for | + | ** Double digests of M55-M66 with EcoRI/PstI. This should generate the following bands: ~2.2 kb for ''P<sub>zntA</sub>'' (M55-M60), ~1.1 kb for rbs+''crtE'' (M61 to M66), and ~2 kb for the pSB1A2 vector. These were run on '''Gel 29'''. M55-M60 unsuccessful; M61-M66 showed a band at the expected location for rbs+''crtE'' but seemed to indicate that L22 was still ligated into BABEL2 instead of pSB1A2. (Yan, AM)<br /> |
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:::: '''[[Edinburgh/30_July_2008|Next Entry >]]''' | :::: '''[[Edinburgh/30_July_2008|Next Entry >]]''' | ||
Latest revision as of 20:38, 29 October 2008
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Week 7
Tuesday 29 July 08
- Transformations of L20 (pSB1A2+rbs+appY) and L21 (pSB1A2+rbs+crtB), plates 63-66, failed.
- Transformation of L25 (pSB1A2+glgC-mut1,2) to plates 67-68 (Yan)
- Cut out putative cenA and cex (both ~1.5kb) fragments from gel 27 (repeat of Gel 26 but stained with SYBRsafe). Purified DNA from gel fragments. (AM)
- Digests of M49-M54 (pSB1A2+rbs+crtI) with EcoRI alone. This should generate a single 4 kb band (doing an EcoRI/PstI double digest would give a vector and insert band both around 2 kb, which would probably run at the same place, making it hard to tell whether there was an insert). These were run on Gel 28. All showed a single band at the expected place. (CF)
- Minipreps of pSB1A2+PzntA (M55-M60) from L23, pSB1A2+rbs+crtE (M61-M66) from L22 and pSB1A2+rbs+crtB (M67) from L21. (Yan, Nimisha)
- Double digests of M55-M66 with EcoRI/PstI. This should generate the following bands: ~2.2 kb for PzntA (M55-M60), ~1.1 kb for rbs+crtE (M61 to M66), and ~2 kb for the pSB1A2 vector. These were run on Gel 29. M55-M60 unsuccessful; M61-M66 showed a band at the expected location for rbs+crtE but seemed to indicate that L22 was still ligated into BABEL2 instead of pSB1A2. (Yan, AM)
- Double digests of M55-M66 with EcoRI/PstI. This should generate the following bands: ~2.2 kb for PzntA (M55-M60), ~1.1 kb for rbs+crtE (M61 to M66), and ~2 kb for the pSB1A2 vector. These were run on Gel 29. M55-M60 unsuccessful; M61-M66 showed a band at the expected location for rbs+crtE but seemed to indicate that L22 was still ligated into BABEL2 instead of pSB1A2. (Yan, AM)
