Team:Stockholm/13 September 2010

From 2010.igem.org

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(New page: {{Stockholm/Top2}} ==Andreas==)
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{{Stockholm/Top2}}
{{Stockholm/Top2}}
==Andreas==
==Andreas==
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===Preparation of Top10 chemically competent cells===
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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.
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Procedures according to protocol. Growth conditions changed to 25 &deg;C, 220 rpm (OD<sub>600</sub> reached after &asymp;6 h).
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A 100 &mu;l aliquot was divided and spread onto Amp 100, Cm 25 and Km 50 plates to verify that they were not contaminated.
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===Cloning of N-CPPs into pSB1C3===
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Sequencing results from 8/9 returned.
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*pSB1C3.nCPP 3 (failed)
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*pSB1C3.nCPP 4 ([[media:PSB1C3.nCPP_4_premix.txt|fasta]])
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*pSB1C3.nCPP 6 ([[media:PSB1C3.nCPP*_6_premix.txt|fasta]])
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*pSB1C3.nCPP 7 ([[media:PSB1C3.nCPP*_7_premix.txt|fasta]])
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*pSB1C3.nCPP 8 ([[media:PSB1C3.nCPP*_8_premix.txt|fasta]])
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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


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).