Team:Chiba/jk/γ/Trl

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実験ログ
出力班


Contents

Time Responce Liquid

purpose

インダクションをかけてからいち早く、確認できる出力を見つけること
To find the earliest output gene after induction

装置&試薬(apparatus&reagents)

装置（apparatus)

  • しんとう培養器(shaking incubator)(37℃,30℃)
  • 48 well plate(deep well)
  • Fluoroskan Ascent 2.5(program:Ascent Software Version 2.6)

試薬(reagents)

  • AHL(100uM,5uM,100nM)
  • IPTG(100nM)
  • X-gal
  • M9
  • PBS

Protocol

  • pre培養(Day,1:pre culture)
  • LB培養液(2ml)に固体培地から大腸菌をpickし３７℃で12時間培養(48穴Deep Well)
  • pick colony(E.coli) from solid culture medium
  • into LB culture fluid(2 ml)
  • cultivate for 12hours in shaking incubator(37℃)
  • 本培(Day,2:main culture)
  • measure OD values and dilution with LB culture fluid to 0.01(OD values)
  • cultivate to 1.0(OD values)
  • wash with physiological saline (2.0min 8,000rpm)x2
  • culture in M9 or PBS and add IPTG or AHL
  • measure OD values ,fluorecent intensity
  • judge whether we can see the color
ODを計測し1/200に希釈しOD=0.01から1.0になるまで培養
OD=1.0になったら生理食塩水でwash(2.0min 8,000rpm)x2
M9またはPBSで培養し、IPTG,AHLを加え計測する。
  • 計測(measurements)

37°Cでしんとう培養
96 shallow well plateに100μLずつ分注し、蛍光強度を測定する。
2h後、3h後…に、100μLずつ分注し、蛍光強度を測定する。
Beam width:Normal Beam
Wavelength pair = 485nm(excitation) and 527nm(emission)

Incubate testing cultures with shaking at 37°C.
After some intervals,aliqupte 100μL of testing cultures into a 96-well plate(shallow well).
Measure fluorescence intensity.
conditions
Beam width:Normal Beam
Wavelength pair = 485nm(excitation) and 527nm(emission)
Retrieved from "https://2008.igem.org/Team:Chiba/protocol/phenotype/timedelay/j"


Result

ホーム メンバー紹介 プロジェクト紹介 Parts Submitted to the Registry モデリング ノート
Retrieved from "http://2008.igem.org/Team:Chiba/jk/%CE%B3/Trl"