Team:Warsaw/Calendar-Main/22 May 2008

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<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPM-T5-AID</a> + 0.01% L-Ara<br>
<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pMPM-T5-AID>pMPM-T5-AID</a> + 0.01% L-Ara<br>
<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a> + 0.5 mM IPTG<br>
<a href=https://2008.igem.org/Wiki/Team:Warsaw/vectors/pTrc99A-AID>pTrc99a-AID</a> + 0.5 mM IPTG<br>
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<a name="fig1"><img src="https://static.igem.org/mediawiki/2008/e/e5/Pcr_fusion_WAW.jpg" width=300/></a>
 
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<var><b>Fig. 1.</b> 1-DNA ladder; 2-PCR product-translational fusion</var></html>
 

Revision as of 11:43, 19 October 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

Preparation of pMPMT5-AID+T7 and pMPMT5+AID-T7

Piotr

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

Michał K.

  1. Isolation of hypothetical pMPM-T5 with transcriptional fusion.
  2. Isolation of hypothetical pMPM-T5 with translational fusion
  3. Control digest of pMPM-T5 with transcriptional fusion with EcoRI (EcoRI buffer).
  4. Control digest of pMPM-T5 with translational fusion with BamHI (BamHI buffer).
  5. Gel electrophoresis (no proper colonies found). Fig. 1.
    6. 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.

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