Team:EPF-Lausanne/Meetings

(Difference between revisions)

Revision as of 10:49, 31 July 2012

Just an sample entry...

For checking PHY42 in a gel, are 900 and 5000 bp OK?
Sequence the PHY42 plasmid (look for standard primers)
List our enzymes
What is the tolerable size of digestion results (for melanopsin, we would have ~4500 nucleotides and ~900 + <size of melanopsin> nucleotides)?
Do we need a backbone (pGL) Maxiprep as well?
How do you check that the backbone has not religated on luciferase? How to remove luciferase? PCR on the digested backbone?
Gel on our pGL4.30 miniprep (for example, digest with HindIII and MfeI, gives ~4000 and ~2000 bp products)
Site-directed mutagenesis on LovTAP (remove the XbaI and EcoRI sites)

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 * Sequence the PHY42 plasmid (look for standard primers)
 * List our enzymes
+* What is the tolerable size of digestion results (for melanopsin, we would have ~4500 nucleotides and ~900 + <size of melanopsin> nucleotides)?
+* Do we need a backbone (pGL) Maxiprep as well?
+* How do you check that the backbone has not religated on luciferase? How to remove luciferase? PCR on the digested backbone?
+* Gel on our pGL4.30 miniprep (for example, digest with HindIII and MfeI, gives ~4000 and ~2000 bp products)
+* Site-directed mutagenesis on LovTAP (remove the XbaI and EcoRI sites)
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