Team:Warsaw/Calendar-Main/9 May 2008

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<var>Fig. 1. 1-DNA ladder; 2-pTRC99a-AID digested with NcoI and HindIII</var>
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<var><b>Fig. 1.</b> 1-DNA ladder; 2-pTRC99a-AID digested with NcoI and HindIII</var>
<img src="https://static.igem.org/mediawiki/2008/0/0f/Gel_out_pmpm_WAW.jpg" width=300/>
<img src="https://static.igem.org/mediawiki/2008/0/0f/Gel_out_pmpm_WAW.jpg" width=300/>
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<var>Fig. 2. 1-DNA ladder; 2-pMPM-T5 digested with NcoI and HindIII</var>
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<var><b>Fig. 2.</b> 1-DNA ladder; 2-pMPM-T5 digested with NcoI and HindIII</var>
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Preparation of pMPMT5+AID construct

Michał K.

  1. Restriction digest of pTrc99A-AID with HindIII and NcoI (1x Tango buffer).
  2. DNA electrophoresis and isolation of DNA encoding AID from proper band (600 bp). Fig. 1.
  3. Restriction digest of pMPM-T5 with HindIII and NcoI (1x Tango buffer), dephosphorylation (CIAP).
  4. DNA electrophoresis and isolation of pMPM-T5 vector from proper band (4250 bp). Fig. 2.
  5. Electrophoresis to estimate the concentration of isolated DNA.
  6. Ligation of isolated DNA fragments.

Fig. 1. 1-DNA ladder; 2-pTRC99a-AID digested with NcoI and HindIII Fig. 2. 1-DNA ladder; 2-pMPM-T5 digested with NcoI and HindIII






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