Team:Warsaw/Calendar-Main/22 May 2008

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Rifampicin Test

Piotr

Overnight inoculation and induction:
Top10
Top10 + 0.5 mM IPTG
pMPM-T5-AID + Glc
pTrc99a-AID + Glc
pMPM-T5-AID + 0.01% L-Ara
pTrc99a-AID + 0.5 mM IPTG
Fig. 1. 1-DNA ladder; 2-PCR product-translational fusion


Preparation of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPMT5-AID+T7</a> and <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPMT5+AID-T7</a>

Piotr

Design of primers for sequencing something we want from pMPM-T5.
Primers:


Michał K.

  1. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPM-T5 with transcriptional fusion</a>.
  2. <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#plasmid_DNA_isolation">Isolation</a> of hypothetical <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPM-T5 with translational fusion</a..
  3. Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAID%2BT7>pMPM-T5 with transcriptional fusion</a> with EcoRI (EcoRI buffer).
  4. Control <a href="https://2008.igem.org/wiki/index.php?title=Wiki/Team:Warsaw/protocols#digest">digest</a> of <a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5%2BAIDT7>pMPM-T5 with translational fusion</a> with BamHI (BamHI buffer).
  5. Gel electrophoresis (no proper colonies found). <a href="https://2008.igem.org/Team:Warsaw/Calendar-Main/22_May_2008#fig1">Fig. 1</a>.

    6. <a name="fig1"><img src="Aid_fusion_digests_WAW.jpg"width=300/></a> Fig. 1. 1-DNA ladder;
    2 and 3-hypothetical pMPM-T5 with transcriptional fusion digested with EcoRI (2 clones);
    4-pMPM-T5-AID digested with EcoRI;
    5-hypothetical pMPM with translational fusion digested with BamHI;
    6-pMPM-T5-AID digested with BamHI.     </html>

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