Team:HokkaidoU Japan/Notebook/Week 3

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Contents

July 16th

Digestion

Results of digestion in 15th.

Digestion result image

Product:Ag43(K346007)=3120bp and 2070bp, pT7-RBS on pSB1K3=2247bp We confirmed there are some kind of restriction enzyme site in K346007 (digested with SpeI, PstI) and pT7-RBS on pSB1K3 was successfully digested with EcoRI and PstI.　Balance between d+(EcoRI) and d+(E&P:cut with EcoRI and PstI) is about 80bp.


Gel Extraction

Gel Extraction for digestion products. We used FastGene Gel&PCR extraction kit(NipponGenetics). Got 50ul of DNA solution.


Ethanol Precipitation

Ethanol Precipitation for digestion and gel extraction products.

  1. Added 5ul of NaoAc, 1.5ul of glycogen and 125ul of 100% ethanol.
  2. Centrifuged in 15000rpm, 10min at 4C.
  3. Remove supernatant and added 220ul of 70% ethanol.
  4. Centrifuged in 15000rpm, 10min at 4C.
  5. Remove supernatant and air drying in room temperature then added 5ul of DW.



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