Team:UGent/Labjournal
From 2013.igem.org
(Difference between revisions)
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<li> Experiment 1 </li> | <li> Experiment 1 </li> | ||
<ul class="a"> | <ul class="a"> | ||
- | <li> Inoculate <i> E.Coli </i>DH5a + pTGD-ccdA-Pmb1GFP-CmFRT </li> | + | <li> <b>Inoculate</b> <i> E.Coli </i>DH5a + pTGD-ccdA-Pmb1GFP-CmFRT </li> |
- | <li> Purify plasmid pTGD-ccdA-Pmb1GFP-CmFRT using Qiagen spin mini kit: Nanodrop -- <b>362.4 ng/µL</b> </li> | + | <li> <b>Purify plasmid pTGD-ccdA-Pmb1GFP-CmFRT</b> using Qiagen spin mini kit: Nanodrop -- <b>362.4 ng/µL</b> </li> |
- | <li> Preparative Restriction Digest (RD) of purified plasmid: BspHI & BstAPI: expected fragments of 5541bp and 903 bp </li> | + | <li> <b>Preparative Restriction Digest (RD)</b> of purified plasmid: BspHI & BstAPI: expected fragments of 5541bp and 903 bp </li> |
- | <li> Gel purify RD-fragment of 5541 bp using Qiagen Qiaquick gel extraction kit: Nanodrop -- 70.4 ng/µL</li> | + | <li> <b>Gel purify RD-fragment of 5541 bp</b> using Qiagen Qiaquick gel extraction kit: Nanodrop -- <b>70.4 ng/µL</b></li> |
- | <li> HiFi PCR using Primestar polymerase on RD (HR-ccdA-Pmb1GFP-HR)and on plasmid pTGD-ccdA-PMbAFGP-CmFRT with MDM588 & MDM589. Analytical gel: <b>negative </b></li> | + | <li> <b>HiFi PCR using Primestar polymerase</b> on RD (HR-ccdA-Pmb1GFP-HR)and on plasmid pTGD-ccdA-PMbAFGP-CmFRT with MDM588 & MDM589. Analytical gel: <b>negative </b></li> |
- | <li> PCR using Q5 polymerase on RD (HR-ccdA-Pmb1GFP-HR)and on plasmid pTGD-ccdA-PMbAFGP-CmFRT with MDM588 & MDM589. Analytical gel: <b> negative </b></li> | + | <li> <b>PCR using Q5 polymerase</b> on RD (HR-ccdA-Pmb1GFP-HR)and on plasmid pTGD-ccdA-PMbAFGP-CmFRT with MDM588 & MDM589. Analytical gel: <b> negative </b></li> |
- | <li> PCR using Roche on RD (HR-ccdA-Pmb1GFP-HR)and on plasmid pTGD-ccdA-PMbAFGP-CmFRT with MDM588 & MDM589. Analytical gel: <b> negative </b></li> | + | <li> <b>PCR using Roche</b> on RD (HR-ccdA-Pmb1GFP-HR)and on plasmid pTGD-ccdA-PMbAFGP-CmFRT with MDM588 & MDM589. Analytical gel: <b> negative </b></li> |
- | <li> Touchdown PCR using Q5 polymerase on RD (HR-ccdA-Pmb1GFP-HR)and on plasmid pTGD-ccdA-PMbAFGP-CmFRT with MDM588 & MDM589. Analytical gel: <b> negative </b></li> | + | <li> <b>Touchdown PCR using Q5 polymerase</b> on RD (HR-ccdA-Pmb1GFP-HR)and on plasmid pTGD-ccdA-PMbAFGP-CmFRT with MDM588 & MDM589. Analytical gel: <b> negative </b></li> |
<li> Control plasmid pTGD-ccdA-PMbAFGP-CmFRT: RD with AclI. Expected fragments:.............. Analytical gel: <b> negative </b> --> <u> Problem with plasmid pTGD-ccdA-PMbAFGP-CmFRT </u></li> | <li> Control plasmid pTGD-ccdA-PMbAFGP-CmFRT: RD with AclI. Expected fragments:.............. Analytical gel: <b> negative </b> --> <u> Problem with plasmid pTGD-ccdA-PMbAFGP-CmFRT </u></li> | ||
<li> <b> Transformation</b>: Knock in with lineair DNA by electroporation. Incubation of the transformed cells and transfer the culture on a Cm plate. </li> | <li> <b> Transformation</b>: Knock in with lineair DNA by electroporation. Incubation of the transformed cells and transfer the culture on a Cm plate. </li> |
Revision as of 17:14, 27 August 2013
Tweets van @iGEM_UGent |
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