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Team:UC Chile2/Cyanolux/Results
From 2012.igem.org
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Our first attempts to construct plasmid pSB1C3_IntK was through a Gibson Assembly reaction of 6 different parts (see construct detail [[http://partsregistry.org/Part:BBa_K743004| BBa_K743004]]) all at once, however we were unsuccessful due to many reasons, many which became clear afterwards (see NOTEPAD WEEK SOMETHING ABOUT GIBSON). We did many attempts betting on depletion of PpsbAB (140bp) due to T5 exonuclease where we changed concentration ratios of the parts, nevertheless we were not able to assemble the parts. | Our first attempts to construct plasmid pSB1C3_IntK was through a Gibson Assembly reaction of 6 different parts (see construct detail [[http://partsregistry.org/Part:BBa_K743004| BBa_K743004]]) all at once, however we were unsuccessful due to many reasons, many which became clear afterwards (see NOTEPAD WEEK SOMETHING ABOUT GIBSON). We did many attempts betting on depletion of PpsbAB (140bp) due to T5 exonuclease where we changed concentration ratios of the parts, nevertheless we were not able to assemble the parts. | ||
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| - | During late June we were able to Biobrick the recombination sites ([[http://partsregistry.org/Part:BBa_K743000 | Part:BBa_K743000]] and [[http://partsregistry.org/Part:BBa_K743001 | Part:BBa_K743001]]) and we decided to use standard assembly to construct a smaller version of pSB1C3_IntK | + | |
| + | During late June we were able to Biobrick the recombination sites ([[http://partsregistry.org/Part:BBa_K743000 | Part:BBa_K743000]] and [[http://partsregistry.org/Part:BBa_K743001 | Part:BBa_K743001]]) and we decided to use standard assembly to construct a smaller version of pSB1C3_IntK [[http://partsregistry.org/Part:BBa_K743006| K743006]] | ||
<h2>Assembling DNA parts</h2> | <h2>Assembling DNA parts</h2> | ||
Revision as of 15:30, 25 September 2012
Question:
Should we separate plasmids from Biobricks? Should each section apply to each construct?
Contents |
Construction of plasmids
Our first attempts to construct plasmid pSB1C3_IntK was through a Gibson Assembly reaction of 6 different parts (see construct detail BBa_K743004) all at once, however we were unsuccessful due to many reasons, many which became clear afterwards (see NOTEPAD WEEK SOMETHING ABOUT GIBSON). We did many attempts betting on depletion of PpsbAB (140bp) due to T5 exonuclease where we changed concentration ratios of the parts, nevertheless we were not able to assemble the parts.
During late June we were able to Biobrick the recombination sites ( Part:BBa_K743000 and Part:BBa_K743001) and we decided to use standard assembly to construct a smaller version of pSB1C3_IntK K743006
Assembling DNA parts
Confirming
Corroborating sequences
Conclusions
Transforming Synechocystis PCC 6803
Transformation
Reestreaking
Confirming DNA
Conclusions
Revealing phenotypes
Microscopy
Luminometer readings
Conclusions
