Team:Wageningen UR/Journal/week9

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week 9: 25 june - 1 july

Office work

Lab work

General

Cloning of BBa_J04500 (IPTG inducible promotor with RBS) and BBa_K197022/38 (Berkeley KILR coil)

  • BBa_J04500 was solubilized from the Standard registry parts plate I with 10µl MQ water
  • BBa_K197022/38 was ordered from the registry
  • transformation of E.coli with both plasmids
  • 5 samples (due to variation of transformation protocol)
  • plated on LB-agar with corresponding antibiotic
  • 5 colonies picked per plate and grown in 5 ml liquid LB overnight
  • subsequent miniprep and digestion check
  • electrophoresis gel showed the expected bands after 20 min running, unfortunately DNA got bleached later by UV light

3 glycerol stocks (25%) of samples 1,3 and 5 were made to test the protocol for freezing cells at 80°C and subsequent thawing and streaking out, which was done successfully the next day.

TuYV

Wednesday:

  • Last week's PCR products(1,1H,2,2H,3,3H) were digested, ligated into the linearized pSB1A3 backbone and transformed into our electrocompetent DH5-alpha cells.

Then after transformation, we checked the colonies on the plate, but only few colonies on positive control and sample 1(TuYV coat protein), 1H(TuYV coat protein his-tag) and 2 plate (TuYV coat protein + 1/2 readthrough).


Friday:

  • Colonies from 1, 1H and 2 plate were picked and checked by colony PCR. But no bands were showed later on the agrose gel check.

Written by: Han Yue


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