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Team:HokkaidoU Japan/Notebook/plastic Week 10
From 2012.igem.org
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==September 5th== | ==September 5th== | ||
<div> | <div> | ||
| + | |||
| + | ===Mini-prep=== | ||
| + | <p> | ||
| + | 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. | ||
| + | </p> | ||
==Colony PCR== | ==Colony PCR== | ||
Revision as of 12:28, 5 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.
