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SDU-Denmark/9 July 2010
From 2010.igem.org
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2) The FlhDC operon entails an internal PST1 site, so we need to induce a silent mutation. For this we need the primers mentioned yesterday. We have fixed the primer annealing temperature problem and have ordered the primers. We hope to get them medio next week.' | 2) The FlhDC operon entails an internal PST1 site, so we need to induce a silent mutation. For this we need the primers mentioned yesterday. We have fixed the primer annealing temperature problem and have ordered the primers. We hope to get them medio next week.' | ||
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These two steps are the most urgent that we work on now. Later on we have to do as follows: | These two steps are the most urgent that we work on now. Later on we have to do as follows: | ||
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3) We need to insert the FlhDC operon into the psb3k3 plasmid. | 3) We need to insert the FlhDC operon into the psb3k3 plasmid. | ||
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''BioBrick Design:'' No Change | ''BioBrick Design:'' No Change | ||
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--[[User:Louch07|Louch07]] 16:00, 9 July 2010 (UTC) | --[[User:Louch07|Louch07]] 16:00, 9 July 2010 (UTC) | ||
Revision as of 14:01, 9 July 2010
Flagella
Overview of the projekt progress and process:
1) We have had problems with out PCR's, none of them work! We have spent the last week trouble-shooting to find the problem and are down to the last few possible causes. It seems now, that the most likely cause is the annealing temperature of the primers, it is too low and the differences between the FW- and the RV-primers are too grate. We hope to solve the mystery soon. When the primer problem is solved we can finally extract the FlhDC operon from the bacteria.
2) The FlhDC operon entails an internal PST1 site, so we need to induce a silent mutation. For this we need the primers mentioned yesterday. We have fixed the primer annealing temperature problem and have ordered the primers. We hope to get them medio next week.'
These two steps are the most urgent that we work on now. Later on we have to do as follows:
3) We need to insert the FlhDC operon into the psb3k3 plasmid.
4) The psb3k3-FlhDC plasmid has to be transformed into cells.
5) Detect if cells are hyperflagellated.
Progress report: Pernille, Sheila and I have worked as a group for the first time today. We have done Miniprep to extract the plasmids we want to insert our biobricks in and Chromosomal DNA PCR to test our primers and the perfect annealing temperature(documented in labnotes)
Working Hypothesis: No Change
BioBrick Design: No Change
--Louch07 16:00, 9 July 2010 (UTC)
