Team:Warsaw/Calendar-Main/1 October 2008

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Preparation of BioBricks

Michał K.

  1. Addition of 5 μl T4 ligase buffer and 2 μl of T4 ligase to digest of pLac_OmpA_omega fragment. Ligation for 1 hr.
  2. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPMT5+AID</a> plasmid using <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDl">AIDl</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#AIDP_HindSpeNotPst">AIDP_HindSpeNotPst</a> primers (annealing temperature 58 °C; elongation length 60s) to obtain AID fragment.


  3. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#pcr">PCR</a> on ligation of pLac_OmpA_omega fragment using <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#PlacL_XNE">PlacL_XNE</a> and <a href="https://2008.igem.org/Wiki/Team:Warsaw/primers#LinP_BS">LinP_BS</a> primers (annealing temperature 58 °C; elongation length 90s) to obtain pLac_OmpA_omega fragment without EcoRI.
  4. Gel electrophoresis of PCR products and <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">gel-out</a> of proper bands (AID - 600 bp and pLac_Omp_omega_ - 1200 bp).
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