"
Team:HokkaidoU Japan/Notebook/plastic Week 10
From 2012.igem.org
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.
