Difference between revisions of "Part:BBa K314110"
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[[Image:Washington_f1_origin_gel.png|thumb|right|300px|'''Figure 1. Single stranded DNA harvested from phage and run on a gel. The higher bands are phage genomic DNA while the lower bands at roughly 2-3kb correspond to the expected plasmid size.''']] | [[Image:Washington_f1_origin_gel.png|thumb|right|300px|'''Figure 1. Single stranded DNA harvested from phage and run on a gel. The higher bands are phage genomic DNA while the lower bands at roughly 2-3kb correspond to the expected plasmid size.''']] | ||
The f1 origin was tested by comparing single strand DNA harvest using part of the [http://2010.igem.org/Team:Washington/Protocols/KunkelCapD Kunkel's mutagenesis] protocol. CJ236 cells were infected with M13K07 helper phage. The CJ236 cells varied in the present or absence of the f1 origin on the pSB1A3 or pSB3K3 plasmid. The SS DNA harvest was then run on a 50ml 1% agarose gel at 90V for 45minutes. As is clearly visible on the gel to the right single strand DNA of the plasmid is only made when the f1 origin is present. | The f1 origin was tested by comparing single strand DNA harvest using part of the [http://2010.igem.org/Team:Washington/Protocols/KunkelCapD Kunkel's mutagenesis] protocol. CJ236 cells were infected with M13K07 helper phage. The CJ236 cells varied in the present or absence of the f1 origin on the pSB1A3 or pSB3K3 plasmid. The SS DNA harvest was then run on a 50ml 1% agarose gel at 90V for 45minutes. As is clearly visible on the gel to the right single strand DNA of the plasmid is only made when the f1 origin is present. | ||
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Revision as of 20:40, 27 October 2010
f1 origin
Phage origin recognized by M13. Used to make SS DNA when infected with M13k07 helper phage.
Usage and Biology
Origin of replication of M13 phages.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 126
- 1000COMPATIBLE WITH RFC[1000]
Functional Parameters
The f1 origin was tested by comparing single strand DNA harvest using part of the [http://2010.igem.org/Team:Washington/Protocols/KunkelCapD Kunkel's mutagenesis] protocol. CJ236 cells were infected with M13K07 helper phage. The CJ236 cells varied in the present or absence of the f1 origin on the pSB1A3 or pSB3K3 plasmid. The SS DNA harvest was then run on a 50ml 1% agarose gel at 90V for 45minutes. As is clearly visible on the gel to the right single strand DNA of the plasmid is only made when the f1 origin is present.