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Team:HokkaidoU Japan/Notebook/plastic Week 10
From 2012.igem.org
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| - | + | ==Mini-prep== | |
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Mini-prep for [RBS-phaC] on pSB1A2 and [RBS-phaA] on pSB1A2 liquid culture products cultivated from 3rd. | Mini-prep for [RBS-phaC] on pSB1A2 and [RBS-phaA] on pSB1A2 liquid culture products cultivated from 3rd. | ||
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</p> | </p> | ||
| - | + | ==Colony PCR== | |
<p> | <p> | ||
We confirmed the length of constructs, [RBS-PhaC-RBS-PhaA on pSB1A2] by colony PCR.<br />The result showed some of ligation [RBS-PhaC on pSB1A2] with [RBS-PhaA] went well. | We confirmed the length of constructs, [RBS-PhaC-RBS-PhaA on pSB1A2] by colony PCR.<br />The result showed some of ligation [RBS-PhaC on pSB1A2] with [RBS-PhaA] went well. | ||
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| - | + | ==Gel Extraction== | |
<p> | <p> | ||
Gel Extraction for digestion products. We used FastGene Gel&PCR extraction kit(NipponGenetics). | Gel Extraction for digestion products. We used FastGene Gel&PCR extraction kit(NipponGenetics). | ||
Got 50 ul of DNA solution of phaB. | Got 50 ul of DNA solution of phaB. | ||
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| + | ==liquid culture== | ||
| + | <p> | ||
| + | The media of pGEM phaCAB was removed to polymer producing media. | ||
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</p> | </p> | ||
Revision as of 05:11, 6 September 2012
Contents |
September 3rd
Single colony isolation
The colony of pGEM PhaCAB in JM109 was isolated to plate1 and plate2.
The colony of pGEM PhaCAB in DH5a was isolated to plate3.
Plate 1~3 was started to incubate from 18:00.
Colony PCR
We confirmed the length of constructs, [PhaB on pSB1C3] and [RBS-PhaC-RBS-PhaA on pSB1A2] by colony PCR.
The result showed ligation to take PhaB on pSB1C3 went well.
But ligation [RBS-PhaC on pSB1A2] with [RBS-PhaA] failed.
Liquid culture
We add 2ml LB and 2ul antibiotic to ideal colony suspension and begun to cultivate at 37C, 180rpm.
September 4th
September 5th
Mini-prep
Mini-prep for [RBS-phaC] on pSB1A2 and [RBS-phaA] on pSB1A2 liquid culture products cultivated from 3rd. We used FastGene Plasmid Mini Kit(Nippon Genetics) and got 50 ul of DNA solutions.
Colony PCR
We confirmed the length of constructs, [RBS-PhaC-RBS-PhaA on pSB1A2] by colony PCR.
The result showed some of ligation [RBS-PhaC on pSB1A2] with [RBS-PhaA] went well.
Gel Extraction
Gel Extraction for digestion products. We used FastGene Gel&PCR extraction kit(NipponGenetics). Got 50 ul of DNA solution of phaB.
liquid culture
The media of pGEM phaCAB was removed to polymer producing media.
