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Team:Stockholm/13 September 2010
From 2010.igem.org
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| + | ===Preparation of Top10 chemically competent cells=== | ||
| + | None of the clones streaked onto the Amp 100 plate (11/9) grew, indicating no AmpR contamination in cells. One of the ON cultures were therefore selected for preparation of competent cells. | ||
| + | |||
| + | Procedures according to protocol. Growth conditions changed to 25 °C, 220 rpm (OD<sub>600</sub> reached after ≈6 h). | ||
| + | |||
| + | A 100 μl aliquot was divided and spread onto Amp 100, Cm 25 and Km 50 plates to verify that they were not contaminated. | ||
| + | |||
| + | ===Cloning of N-CPPs into pSB1C3=== | ||
| + | Sequencing results from 8/9 returned. | ||
| + | *pSB1C3.nCPP 3 (failed) | ||
| + | *pSB1C3.nCPP 4 ([[media:PSB1C3.nCPP_4_premix.txt|fasta]]) | ||
| + | *pSB1C3.nCPP 6 ([[media:PSB1C3.nCPP*_6_premix.txt|fasta]]) | ||
| + | *pSB1C3.nCPP 7 ([[media:PSB1C3.nCPP*_7_premix.txt|fasta]]) | ||
| + | *pSB1C3.nCPP 8 ([[media:PSB1C3.nCPP*_8_premix.txt|fasta]]) | ||
| + | |||
| + | Blastn alignments against [[media:Blastn_N-Tra10_pSB1C3.nCPP_11sep.txt|N-Tra10]], [[media:Blastn_N-TAT_pSB1C3.nCPP_11sep.txt|N-TAT]] and [[media:Blastn_N-LMWP_pSB1C3.nCPP_11sep.txt|N-LMWP]] indicated successful cloning of N-Tra10 (clone 7) and N-TAT (clone 4). | ||
Revision as of 17:55, 14 September 2010
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Andreas
Preparation of Top10 chemically competent cells
None of the clones streaked onto the Amp 100 plate (11/9) grew, indicating no AmpR contamination in cells. One of the ON cultures were therefore selected for preparation of competent cells.
Procedures according to protocol. Growth conditions changed to 25 °C, 220 rpm (OD600 reached after ≈6 h).
A 100 μl aliquot was divided and spread onto Amp 100, Cm 25 and Km 50 plates to verify that they were not contaminated.
Cloning of N-CPPs into pSB1C3
Sequencing results from 8/9 returned.
- pSB1C3.nCPP 3 (failed)
- pSB1C3.nCPP 4 (fasta)
- pSB1C3.nCPP 6 (fasta)
- pSB1C3.nCPP 7 (fasta)
- pSB1C3.nCPP 8 (fasta)
Blastn alignments against N-Tra10, N-TAT and N-LMWP indicated successful cloning of N-Tra10 (clone 7) and N-TAT (clone 4).
