Team:Warsaw/Calendar-Main/12 May 2008

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<p>Michał K.:</p>
<p>Michał K.:</p>
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<p>1. Restriction <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digestation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99A-AID</a> with HindIII and NcoI (1x Tango buffer).</p>
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<p>1. Restriction <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digestion</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99A-AID</a> with HindIII and NcoI (1x Tango buffer).</p>
<p>2. DNA electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation of DNA</a> encoding AID from proper band (600 bp).</p>
<p>2. DNA electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation of DNA</a> encoding AID from proper band (600 bp).</p>
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<p>3. Restriction <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digestation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with HindIII and NcoI (1x Tango buffer), <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">dephosphorylation</a> (CIAP).</p>
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<p>3. Restriction <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digestion</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> with HindIII and NcoI (1x Tango buffer), <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#removing">dephosphorylation</a> (CIAP).</p>
<p>4. DNA electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> vector from proper band (4250 bp).</p>
<p>4. DNA electrophoresis and <a href="https://2008.igem.org/Wiki/Team:Warsaw/protocols#DNA_isolation_from_agarose_gel">isolation</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-omega>pMPM-T5</a> vector from proper band (4250 bp).</p>

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Michał K.:

1. Restriction digestion of pTrc99A-AID with HindIII and NcoI (1x Tango buffer).

2. DNA electrophoresis and isolation of DNA encoding AID from proper band (600 bp).

3. Restriction digestion 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).

5. Ligation of isolated DNA fragments.

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