Team:Warsaw/Calendar-Main/2 October 2008

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<h3>Preparation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha</h3>
<h3>Preparation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _alpha</h3>
<h4>Michał K.</h4>
<h4>Michał K.</h4>
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<p> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103009>linker_alpha (BBa_K103009).</p>
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<p> Overnight <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#ligation">ligation</a> of isolated DNA fragments: <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + <a href=http://partsregistry.org/wiki/index.php?title=Part:BBa_K103009>linker_alpha (BBa_K103009)</a>.</p>
<h3>Preparation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega</h3>
<h3>Preparation of <a href=http://partsregistry.org/Part:pSB2K3>pSB2K3</a> + _omega</h3>

Revision as of 22:42, 26 October 2008

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Preparation of pSB2K3 + _alpha

Michał K.

Overnight ligation of isolated DNA fragments: pSB2K3 + linker_alpha (BBa_K103009).

Preparation of pSB2K3 + _omega

Michał K.

Overnight ligation of isolated DNA fragments: pSB2K3 + _omega.

Preparation of BioBricks

Michał K.

  1. Isolation of plasmids from culture inoculated on previous day (pSB1A3+Z(BBa_K103004)).
  2. Control digest of isolated plasmid with EcoRI and PstI (Orange buffer). Gel electrophoresis - proper clones found.
  3. Digest of pLac_OmpA_omega fragment with EcoRI and BcuI (BamHI buffer).
  4. Gel elctrophoresis and gel-out of proper band - 1200 bp.
  5. Colony PCR with OmpaL_N and OmpaP_link primers on colonies from plates with transformations pSB1A3+OmpA (annealing temperature - 55°C,45 s of elongation step).
  6. Gel electrophoresis.
  7. Confirmed transformant colonies inoculated to liquid LB with ampicillin.
  8. Overnight ligation of isolated DNA fragments: pSB2K3 + _alpha, pSB2K3 + _omega and pACYC177 + OmpA_omega_ (both digested with BamHI and PstI - from 30 September).

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