Team:Wageningen UR/Journal/week9
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'''General''' | '''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_J04500 was solubilized from the Standard registry parts plate I with 10µl MQ water | ||
*BBa_K197022/38 was ordered from the registry | *BBa_K197022/38 was ordered from the registry |
Revision as of 13:47, 24 September 2012
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